Styrenyl derivative compounds for treating ophthalmic diseases and disorders

ABSTRACT

The present invention relates generally to compositions and methods for treating neurodegenerative diseases and disorders, particularly ophthalmic diseases and disorders. Provided herein are styrenyl derivative compounds, including but not limited to stilbene derivative compounds, and compositions comprising these compounds, that are useful for treating and preventing ophthalmic diseases and disorders, including age-related macular degeneration (AMD) and Stargardt&#39;s Disease.

CROSS-REFERENCE

This application is a continuation application of U.S. patentapplication Ser. No. 12/107,040, filed Apr. 21, 2008, which claims thebenefit of U.S. Provisional Application No. 60/937,002, filed Jun. 22,2007, and U.S. Provisional Application No. 60/913,241, filed Apr. 20,2007, the contents of which-are incorporated herein by reference intheir entirety.

BACKGROUND OF THE INVENTION

Neurodegenerative diseases, such as glaucoma, macular degeneration, andAlzheimer's disease, affect millions of patients throughout the world.Because the loss of quality of life associated with these diseases isconsiderable, drug research and development in this area is of greatimportance.

Macular degeneration affects between ten and fifteen million patients inthe United States, and it is the leading cause of blindness in agingpopulations worldwide. Age-related macular degeneration affects centralvision and causes the loss of photoreceptor cells in the central part ofthe retina called the macula. Macular degeneration can be classifiedinto two types: dry-type and wet-type. The dry-form is more common thanthe wet, with about 90% of age-related macular degeneration (AMD)patients diagnosed with the dry-form. The wet-form of the disease andgeographic atrophy, which is the end-stage phenotype of dry AMD, causesthe most serious vision loss. All patients who develop wet-form AMD arebelieved to previously have been developing dry-form AMD for a prolongedperiod of time. The exact causes of age-related macular degeneration arestill unknown. The dry-form of AMD may result from the senescence andthinning of macular tissues associated with the deposition of pigment inthe macular retinal pigment epithelium. In wet AMD, new blood vesselsgrow beneath the retina, form scar tissue, bleed, and leak fluid. Theoverlying retina can be severely damaged, creating “blind” areas in thecentral vision.

For the vast majority of patients who have the dry-form of maculardegeneration, no effective treatment is yet available. Because thedry-form precedes development of the wet-form of macular degeneration,therapeutic intervention to prevent or delay disease progression in thedry-form AMD would benefit patients with dry-form AMD and might reducethe incidence of the wet-form.

Decline of vision noticed by the patient or characteristic featuresdetected by an ophthalmologist during a routine eye exam may be thefirst indicator of age-related macular degeneration. The formation of“drusen,” or membranous debris beneath the retinal pigment epithelium ofthe macula is often the first physical sign that AMD is developing. Latesymptoms include the perceived distortion of straight lines and, inadvanced cases, a dark, blurry area or area with absent vision appearsin the center of vision; and/or there may be color perception changes.

Different forms of genetically-linked macular degenerations may alsooccur in younger patients. In other maculopathies, there are hereditary,nutritional, traumatic, infection, or other ecologic factors.

Glaucoma is a broad term used to describe a group of diseases thatcauses a slowly progressive visual field loss, usually asymptotically.The lack of symptoms may lead to a delayed diagnosis of glaucoma untilthe terminal stages of the disease. The prevalence of glaucoma isestimated to be 2.2 million in the United States, with about 120,000cases of blindness attributable to the condition. The disease isparticularly prevalent in Japan, which has four million reported cases.In many parts of the world, treatment is less accessible than in theUnited States and Japan, thus glaucoma ranks as a leading cause ofblindness worldwide. Even if subjects afflicted with glaucoma do notbecome blind, their vision is often severely impaired.

The progressive loss of peripheral visual field in glaucoma is caused bythe death of ganglion cells in the retina. Ganglion cells are a specifictype of projection neuron that connects the eye to the brain. Glaucomais usually accompanied by an increase in intraocular pressure. Currenttreatment includes use of drugs that lower the intraocular pressure;however, contemporary methods to lower the intraocular pressure areoften insufficient to completely stop disease progression. Ganglioncells are believed to be susceptible to pressure and may sufferpermanent degeneration prior to the lowering of intraocular pressure. Anincreasing number of cases of normal-tension glaucoma are observed inwhich ganglion cells degenerate without an observed increase in theintraocular pressure. Because current glaucoma drugs only treatintraocular pressure, a need exists to identify new therapeutic agentsthat will prevent or reverse the degeneration of ganglion cells.

Recent reports suggest that glaucoma is a neurodegenerative disease,similar to Alzheimer's disease and Parkinson's disease in the brain,except that it specifically affects retinal neurons. The retinal neuronsof the eye originate from diencephalon neurons of the brain. Thoughretinal neurons are often mistakenly thought not to be part of thebrain, retinal cells are key components of the central nervous system,interpreting the signals from the light-sensing cells.

Alzheimer's disease (AD) is the most common form of dementia among theelderly. Dementia is a brain disorder that seriously affects a person'sability to carry out daily activities. Alzheimer's is a disease thataffects four million people in the United States alone. It ischaracterized by a loss of nerve cells in areas of the brain that arevital to memory and other mental functions. Currently available drugscan ameliorate AD symptoms for a relatively period of time, but no drugsare available that treat the disease or completely stop the progressivedecline in mental function. Recent research suggests that glial cellsthat support the neurons or nerve cells may have defects in ADsufferers, but the cause of AD remains unknown. Individuals with AD seemto have a higher incidence of glaucoma and age-related maculardegeneration, indicating that similar pathogenesis may underlie theseneurodegenerative diseases of the eye and brain. (See Giasson et al.,Free Radic. Biol. Med. 32:1264-75 (2002); Johnson et al., Proc. Natl.Acad. Sci. USA 99:11830-35 (2002); Dentchev et al., Mol. Vis. 9:184-90(2003)).

Neuronal cell death underlies the pathology of these diseases.Unfortunately, very few compositions and methods that enhance retinalneuronal cell survival, particularly photoreceptor cell survival, havebeen discovered. A need therefore exists to identify and developcompositions that can be used for treatment and prophylaxis of a numberof retinal diseases and disorders that have neuronal cell death as aprimary, or associated, element in their pathogenesis.

In vertebrate photoreceptor cells, the irradiance of a photon causesisomerization of 11-cis-retinylidene chromophore toall-trans-retinylidene and uncoupling from the visual opsin receptors.This photoisomerization triggers conformational changes of opsins,which, in turn, initiate the biochemical chain of reactions termedphototransduction (Filipek et al., Annu. Rev. Physiol. 65:851-79(2003)). Regeneration of the visual pigments requires that thechromophore be converted back to the 11-cis-configuration in theprocesses collectively called the retinoid (visual) cycle (see, e.g.,McBee et al., Prog. Retin. Eye Res. 20:469-52 (2001)). First, thechromophore is released from the opsin and reduced in the photoreceptorby retinol dehydrogenases. The product, all-trans-retinol, is trapped inthe adjacent retinal pigment epithelium (RPE) in the form of insolublefatty acid esters in subcellular structures known as retinosomes(Imanishi et al., J. Cell Biol. 164:373-87 (2004)).

In Stargardt's disease (Allikmets et al., Nat. Genet. 15:236-46 (1997)),a disease associated with mutations in the ABCR transporter that acts asa flippase, the accumulation of all-trans-retinal may be responsible forthe formation of a lipofuscin pigment, A2E, which is toxic towardsretinal pigment epithelial cells and causes progressive retinaldegeneration and, consequently, loss of vision (Mata et al., Proc. Natl.Acad. Sci. USA 97:7154-59 (2000); Weng et al., Cell 98:13-23 (1999)).Treating patients with an inhibitor of retinol dehydrogenases, 13-cis-RA(Isotretinoin, Accutane®, Roche), has been considered as a therapy thatmight prevent or slow the formation of A2E and might have protectiveproperties to maintain normal vision (Radu et al., Proc. Natl. Acad.Sci. USA 100:4742-47 (2003)). 13-cis-RA has been used to slow thesynthesis of 11-cis-retinal by inhibiting 11-cis-RDH (Law et al.,Biochem. Biophys. Res. Commun. 161:825-9 (1989)), but its use can alsobe associated with significant night blindness. Others have proposedthat 13-cis-RA works to prevent chromophore regeneration by bindingRPE65, a protein essential for the isomerization process in the eye(Gollapalli et al., Proc. Natl. Acad. Sci. USA 101:10030-35 (2004)).Gollapalli et al. reported that 13-cis-RA blocked the formation of A2Eand suggested that this treatment may inhibit lipofuscin accumulationand, thus, delay either the onset of visual loss in Stargardt's diseaseor age-related macular degeneration, which are both associated withretinal pigment-associated lipofuscin accumulation. However, blockingthe retinoid cycle and forming unliganded opsin may result in moresevere consequences and worsening of the patient's prognosis (see, e.g.,Van Hooser et al., J. Biol. Chem. 277:19173-82 (2002); Woodruff et al.,Nat. Genet. 35:158-164 (2003)). Failure of the chromophore to form maylead to progressive retinal degeneration and may produce a phenotypesimilar to Leber Congenital Amaurosis (LCA), is a very rare geneticcondition affecting children shortly after birth.

A need exists in the art for an effective treatment for treatingophthalmic diseases or disorders resulting in ophthalmic disfunctionincluding those described above. In particular, there exists a pressingneed for compositions and methods for treating Stargardt's disease andage-related macular degeneration (AMD) without causing further unwantedside effects such as progressive retinal degeneration, LCA-likeconditions, night blindness, or systemic vitamin A deficiency. A needalso exists in the art for effective treatments for other ophthalmicdiseases and disorders that adversely affect the retina.

SUMMARY OF THE INVENTION

The present invention relates to styrenyl derivative compounds, whichcan be inhibitors of an isomerization step of the retinoid cycle and areuseful for treating ophthalmic diseases and disorders. Also provided arepharmaceutical compositions comprising the styrenyl derivative compoundsand methods for treating various ophthalmic diseases using thesecompounds.

In one embodiment is a compound having a structure of Formula (A):

as an isolated E or Z geometric isomer or a mixture of E and Z geometricisomers, as a tautomer or a mixture of tautomers, as a stereoisomer oras a pharmaceutically acceptable salt, hydrate, solvate, N-oxide orprodrug thereof, wherein: R1 and R2 are each the same or different andindependently hydrogen or alkyl; R3, R4, R5 and R6 are each the same ordifferent and independently hydrogen, halogen, nitro, —NH2, —NHR13,—N(R13)2, —OR12, alkyl or fluoroalkyl;R7 and R8 are each the same or different and independently hydrogen oralkyl; or R7 and R8 together with the carbon atom to which they areattached, form a carbocyclyl or heterocyclyl; or R7 and R8 together forman imino;R9 is hydrogen, alkyl, carbocyclyl, heterocyclyl, —C(═O)R13, —SO2R13,—CO2R13, —CONH2, —CON(R13)2 or —CON(H)R13;R10 is hydrogen or alkyl; or R9 and R10, together with the nitrogen atomto which they are attached, form an N-heterocyclyl;R11 is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;each R12 is independently selected from hydrogen or alkyl; each R13 isindependently selected from alkyl, carbocyclyl, heterocyclyl, aryl orheteroaryl;Z is a bond, Y or W—Y, wherein W is —C(R14)(R15)-, —O—, —S—, —S(═O)—,—S(═O)2- or —N(R12)-; Y is —C(R16)(R17)- or —C(R16)(R17)-C(R21)(R22)-;R14 and R15 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19, carbocyclyl orheterocyclyl; or R14 and R15 together form an oxo, an imino, an oximo,or a hydrazino; R16 and R17 are each the same or different andindependently hydrogen, halogen, alkyl, fluoroalkyl, —OR12, —NR18R19,carbocyclyl or heterocyclyl; or R16 and R17 together form an oxo; oroptionally, R14 and R16 together form a direct bond to provide a doublebond connecting W and Y; or optionally, R14 and R16 together form adirect bond, and R15 and R17 together form a direct bond to provide atriple bond connecting W and Y; each R18 and R19 is independentlyselected from hydrogen, alkyl, carbocyclyl, or —C(═O)R13, —SO2R13,—CO2R13, —CONH2, —CON(R13)2 or —CON(H)R13; or R18 and R19, together withthe nitrogen atom to which they are attached, form an N-heterocyclyl;R21 and R22 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19, carbocyclyl orheterocyclyl;provided that when R11 is phenyl, the compound of Formula (A) is not:

-   2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]acetamide;-   (2S,3R)-amino-3-hydroxy-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)-ethenyl]phenyl]-butanamide;-   L-glutamic acid,    1-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]ester;-   glycine, 3-hydroxy-5-[(1E)-2-(4-hydroxyphenyl)ethenyl]phenyl ester;-   (2S)-2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]propanamide;-   (2S)-2-amino-3-hydroxy-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]propanamide;-   (2S)-2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]-4-methyl-pentanamide;-   (2S)-2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]-3-methyl-butanamide;    or-   2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenylbutanamide.

In another embodiment is the compound wherein each of R1 and R8 ishydrogen, and the compound has a structure of Formula (B):

whereinR2 is H or C1-C6 alkyl;each of R3, R4, R5, and R6 is the same or different and independentlyhydrogen, halo, C1-C6 alkyl, or —OR12;R7 is H or C1-C6 alkyl;R9 is hydrogen, C1-C6 alkyl, —(CH2)nOH wherein n is 2-6, or —C(═O)R13;R10 is hydrogen or C1-C6 alkyl; or R9 and R10, together with thenitrogen atom to which they are attached, form an N-heterocyclyl;W is —C(R14)(R15)-, —O—, —S—, —S(═O)—, —S(═O)2- or —N(H)—;Y is —C(R16)(R17)-; each R12 is independently hydrogen or C1-C6 alkyl;each R13 is independently a C1-C6 alkyl; R14 and R15 are each the sameor different and independently selected from hydrogen, halogen, C1-C6alkyl, fluoroalkyl, —OR12, or —NH2; or R14 and R15 together form an oxo,an imino, an oximo, or a hydrazino; R16 and R17 are each the same ordifferent and independently hydrogen, halogen, C1-C6 alkyl, or —OR12; orR16 and R17 together form an oxo; or optionally, R14 and R16 togetherform a direct bond to provide a double bond connecting W and Y; oroptionally, R14 and R16 together form a direct bond, and R15 and R17together form a direct bond to provide a triple bond connecting W and Y;and R11 is selected from:a) alkyl;b) phenyl substituted with alkyl, —OR12, —O(CH2)mOCH3 wherein m is 1-6,alkenyl, alkynyl, halogen, fluoroalkyl, phenyl, —SCH3, or aralkyl;c) naphthenyl optionally substituted with alkyl, halogen, or —OR12;d) carbocyclyl; ore) cyclohexenyl optionally substituted with alkyl,provided that R11 is not 3,4,5-tri-methoxyphenyl.In a further embodiment is the compound of Formula (A) or (B) wherein R2is hydrogen or n-butyl. In an additional embodiment is the compound ofFormula (A) or (B) wherein halogen is fluoro or chloro.

In yet another embodiment is the compound of either Formula (A) or (B)wherein each of R3, R4, R5, and R6 is the same or different andindependently hydrogen, halogen, methyl, or methoxy.

In an additional embodiment is the compound of Formula (A) or (B),wherein at least two of R3, R4, R5, and R6 are hydrogen.

In a further embodiment is the compound of Formula (A) or (B), wherein Wis —C(R14)(R15)-, and wherein R14 and R15 are each the same or differentand independently hydrogen, fluoro, methyl, ethyl, trifluoromethyl, —OH,—OCH3, or —NH2.

In another embodiment is the compound of Formula (A) or (B), whereineach of R14 and R15 is hydrogen.

In an additional embodiment is the compound of Formula (A) or (B),wherein Y is —CH2-, —CH(CH3)-, —C(CH3)2-, —C(H)OH—, —C(H)F—, —CF2-, or—C(═O)—.

In yet another embodiment is the compound of Formula (A) or (B), whereinR11 is phenyl substituted with alkyl, —OR12, —O(CH2)nOCH3 wherein n is2-6, alkenyl, alkynyl, halogen, fluoroalkyl, phenyl, or —SCH3.

In one embodiment is the compound of Formula (A) or (B), wherein R11 isnaphthenyl substituted with —OR12, wherein R12 is hydrogen or C1-C6alkyl.

In an additional embodiment is the compound of Formula (A) or (B),wherein R11 is cyclohexenyl optionally substituted with C1-C6 alkyl. Insome embodiments R11 is tri-methyl-cyclohexenyl.

In additional embodiments is the compound of Formula (A) or (B), whereinR11 is alkyl optionally substituted with —OR12 wherein R12 is hydrogenor C1-C6 alkyl.

In an additional embodiment is the compound of Formula (A), wherein R11is aryl or carbocyclyl.

In an alternative embodiment is the compound of Formula (C), wherein R11is aryl:

Formula (C) as an isolated E or Z geometric isomer or a mixture of E andZ geometric isomers, as a tautomer or a mixture of tautomers, as astereoisomer, or as a pharmaceutically acceptable salt, hydrate,solvate, N-oxide or prodrug thereof, wherein:

R1 and R2 are each the same or different and independently hydrogen oralkyl;

R3, R4, R5 and R6 are each the same or different and independentlyhydrogen, halogen, nitro, —NH2, —NHR13, —N(R13)2, —OR12, alkyl orfluoroalkyl;

R7 and R8 are each the same or different and independently hydrogen oralkyl;

R9 is hydrogen, alkyl, carbocyclyl or —C(═O)R13;

R10 is hydrogen or alkyl; or R9 and R10, together with the nitrogen atomto which they are attached, form an N-heterocyclyl;

each R12 is independently selected from hydrogen or alkyl;

R13 is alkyl, carbocyclyl or aryl;

W is —C(R14)(R15)-, —O—, —S—, —S(═O)—, —S(═O)2- or —N(R12)-;

Y is —C(R16)(R17)-;

R14 and R15 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19 or carbocyclyl; or R14 andR15 together form an oxo, an imino, an oximo, or a hydrazino;

R16 and R17 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19 or carbocyclyl; or R16 andR17 together form an oxo;

optionally, R14 and R16 together form a direct bond to provide a doublebond connecting W and Y; or

optionally, R14 and R16 together form a direct bond, and R15 and R17together form a direct bond to provide a triple bond connecting W and Y;

each R18 and R19 is independently selected from hydrogen, alkyl,carbocyclyl, or —C(═O)R13;

t is 0, 1, 2, 3, 4 or 5; and

each R20 is the same or different and independently selected from alkyl,—OR12, —SR12, alkenyl, alkynyl, halo, fluoroalkyl, aryl or aralkyl; ortwo adjacent R20 groups, together with the two carbon atoms to whichthey are attached, form a fused phenyl ring.

In an additional embodiment is the compound wherein each of R9 and R10is hydrogen and the compound has a structure of Formula (D):

In a further embodiment is the compound of Formula (E) wherein W is—C(R14)(R15)-:

as an isolated E or Z geometric isomer or a mixture of E and Z geometricisomers, as a tautomer or a mixture of tautomers, as a stereoisomer, oras a pharmaceutically acceptable salt, hydrate, solvate, N-oxide orprodrug thereof, wherein:R1 and R2 are each the same or different and independently hydrogen oralkyl;R3, R4, R5 and R6 are each the same or different and independentlyhydrogen, halogen, nitro, —NH2, —NHR13, —N(R13)2, —OR12, alkyl orfluoroalkyl;R7 and R8 are each the same or different and independently hydrogen oralkyl;each R12 is independently selected from hydrogen or alkyl;R13 is alkyl, carbocyclyl or aryl;R14 and R15 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19 or carbocyclyl; or R14 andR15 form an oxo, an imino, an oximo, or a hydrazino;R16 and R17 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19 or carbocyclyl; or R16 andR17 form an oxo;each R18 and R19 are each the same or different and independentlyhydrogen, alkyl, carbocyclyl, or —C(═O)R13;t is 0, 1, 2, 3, 4 or 5; andeach R20 is the same or different and independently alkyl, —OR12, —SR12,alkenyl, alkynyl, halo, fluoroalkyl, aryl or aralkyl, or two adjacentR20, together with the two carbon atoms to which they are attached, forma fused phenyl ring.In a additional embodiment is the compound of Formula (E) wherein t is0, 1, 2 or 3;each R20 is independently alkyl, —OR12, —SR12, alkynyl, phenyl, halo orfluoroalkyl; andR3, R4, R5 and R6 are each independently hydrogen, alkyl, —OR12, halo orfluoroalkyl.

In a further embodiment is the compound of Formula (E), wherein R7, R8,R14, R15, R16 and R17 are each independently hydrogen, halogen, alkyl or—OR12, wherein each R12 is independently selected from hydrogen oralkyl.

In an additional embodiment is the compound of Formula (E), wherein:

t is 2 or 3; two adjacent R20, together with the two carbon atoms towhich they are attached, form a fused phenyl ring; and

R3, R4, R5 and R6 are each independently hydrogen, alkyl, halo orfluoroalkyl.

In another embodiment is the compound of Formula (E), wherein R7, R8,R14, R15, R16 and R17 are each independently hydrogen, halogen, alkyl or—OR12.

In an additional embodiment is the compound of Formula (E), wherein W is—O—, —S—, —S(═O)—, —S(═O)2- or —N(R12)-.

In yet another embodiment is the compound of Formula (E), wherein: t is0, 1, 2 or 3; each R20 is independently alkyl, —OR12 or halo; and R3,R4, R5 and R6 are each independently hydrogen, alkyl or halo.

In a specific embodiment is the compound of Formula (E) wherein W and Yare connected by a double bond.

In an additional embodiment is the compound of Formula (E), wherein R9and R10 together with the nitrogen to which they are attached form aN-heterocyclyl.

In an additional embodiment is the compound of Formula (E), wherein theN-heterocyclyl is morpholinyl, pyrrolidinyl, piperidinyl or piperazinyl.

In an additional embodiment is the compound of Formula (E), wherein:each of R1 and R2 is hydrogen; t is 0, 1, 2 or 3; each R20 isindependently alkyl or halo; and R3, R4, R5 and R6 are eachindependently hydrogen, alkyl or halo.

In another embodiment is the compound of Formula (E), wherein W is—C(R14)(R15)-.

In an additional embodiment is the compound of Formula (E), wherein R9is alkyl or —C(═O)R13, wherein R13 is alkyl, and R10 is hydrogen oralkyl.

In an additional embodiment is the compound of Formula (E), wherein:each of R1 and R2 is hydrogen; t is 0, 1, 2 or 3; each R20 isindependently alkyl or halo; and R3, R4, R5 and R6 are eachindependently hydrogen, alkyl or halo.

In an additional embodiment is the compound of Formula (E), wherein W is—C(R14)(R15)-.

In an additional embodiment is the compound of Formula (A), wherein R11is 3-, 4-, 5-, 6-, 7- or 8-member cycloalkyl. In some embodiments R11 iscyclohexyl.

In an additional embodiment is the compound of Formula (A), wherein R11is 5-, 6-, or 7-member cycloalkenyl.

In an additional embodiment is the compound of Formula (F), wherein R11is cyclohexenyl:

as an isolated E or Z geometric isomer or a mixture of E and Z geometricisomers, as a tautomer or a mixture of tautomers, as a stereoisomer, oras a pharmaceutically acceptable salt, hydrate, solvate, N-oxide orprodrug thereof, wherein:R1 and R2 are each the same or different and independently hydrogen oralkyl;R3, R4, R5 and R6 are each the same or different and independentlyhydrogen, halogen, nitro, —NH2, —NHR13, —N(R13)2, —OR12, alkyl orfluoroalkyl;R7 and R8 are each the same or different and independently hydrogen oralkyl;R9 is hydrogen, alkyl, carbocyclyl or —C(═O)R13;R10 is hydrogen or alkyl; or R9 and R10, together with the nitrogen atomto which they are attached, form an N-heterocyclyl;each R12 is independently selected from hydrogen or alkyl;each R13 is independently selected from alkyl, carbocyclyl or aryl;W is —C(R14)(R15)-, —O—, —S—, —S(═O)—, —S(═O)2- or —N(R12)-;Y is —C(R16)(R17)-;R14 and R15 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19 or carbocyclyl; or R14 andR15 together form an oxo, an imino, an oximo, or a hydrazino;R16 and R17 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19 or carbocyclyl; or R16 andR17 together form an oxo;optionally, R14 and R16 together form a direct bond to provide a doublebond connecting W and Y; or R14 and R16 together form a direct bond, andR15 and R17 together form a direct bond to provide a triple bondconnecting W and Y;each R18 and R19 is independently selected from hydrogen, alkyl,carbocyclyl, or —C(═O)R13;p is 0, 1, 2, 3, 4, 5, 7, 8 or 9; andeach R21 is the same or different and independently selected from alkyl,—OR12, alkenyl, alkynyl, halo, fluoroalkyl or aralkyl.

In an additional embodiment is the compound wherein W is —C(R14)(R15)-,and the compound has a structure of Formula (G):

as an isolated E or Z geometric isomer or a mixture of E and Z geometricisomers, as a tautomer or a mixture of tautomers, as a stereoisomer, oras a pharmaceutically acceptable salt, hydrate, solvate, N-oxide orprodrug thereof, wherein:R1 and R2 are each the same or different and independently hydrogen oralkyl;R3, R4, R5 and R6 are each the same or different and independentlyhydrogen, halogen, nitro, —NH2, —NHR13, —N(R13)2, —OR12, alkyl orfluoroalkyl;R7 and R8 are each the same or different and independently hydrogen oralkyl;R9 is hydrogen, alkyl, carbocyclyl or —C(═O)R13;R10 is hydrogen or alkyl; or R9 and R10, together with the nitrogen atomto which they are attached, form an N-heterocyclyl;each R12 is independently selected from hydrogen or alkyl;each R13 is independently alkyl, carbocyclyl or aryl;R14 and R15 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19 or carbocyclyl; or R14 andR15 together form an oxo, an imino, an oximo, or a hydrazino;R16 and R17 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19 or carbocyclyl; or R16 andR17 together form an oxo;optionally, R14 and R16 together form a direct bond to provide a doublebond connecting W and Y; or R14 and R16 together form a direct bond, andR15 and R17 together form a direct bond to provide a triple bondconnecting W and Y;each R18 and R19 is independently selected from hydrogen, alkyl,carbocyclyl, or —C(═O)R13;p is 0, 1, 2, 3, 4, 5, 7, 8 or 9; andeach R21 is the same or different and independently alkyl, —OR12,alkenyl, alkynyl, halo, fluoroalkyl or aralkyl.

In an additional embodiment is the compound of Formula (G), wherein eachof R9 and R10 is hydrogen.

In an additional embodiment is the compound of Formula (G), wherein:each of R1 and R2 is hydrogen; p is 0, 1, 2 or 3; each R21 isindependently alkyl, halo or fluoroalkyl; and each of R3, R4, R5 and R6is independently hydrogen, alkyl, halo, fluoroalkyl or —OR12.

In an additional embodiment is the compound of Formula (G), wherein: R7,R8, R14, R15, R16 and R17 are each independently hydrogen, halogen,alkyl, fluoroalkyl, —OR12 or —NR18R19, wherein each R12 is independentlyhydrogen or alkyl; and each R18 and R19 are independently hydrogen oralkyl.

In an additional embodiment is the compound of Formula (G), wherein R9is alkyl and R10 is hydrogen.

In an additional embodiment is the compound of Formula (G), wherein:each of R1 and R2 is hydrogen; p is 0, 1, 2 or 3; each R21 isindependently alkyl, halo or fluoroalkyl; and each of R3, R4, R5 and R6is independently hydrogen, alkyl, halo, fluoroalkyl or —OR12.

In an additional embodiment is the compound of Formula (G), wherein R7,R8, R14, R15, R16 and R17 are each independently hydrogen, halogen,alkyl, fluoroalkyl or —OR12, wherein each R12 is independently hydrogenor alkyl.

In an additional embodiment is the compound of Formula (G), wherein: R7,R8, R16 and R17 are each independently hydrogen, halogen, alkyl,fluoroalkyl or —OR12, wherein R12 is hydrogen or alkyl; and R14 and R15together form oxo.

In an additional embodiment is the compound of Formula (G), wherein W is—NH— or —O—.

In an additional embodiment is the compound of Formula (G), wherein eachof R1, R2, R9 and R10 is hydrogen.

In an additional embodiment is the compound of Formula (G), wherein: pis 0, 1, 2 or 3; each R21 is independently alkyl or halo; and R3, R4, R5and R6 are each independently hydrogen, alkyl, halo or fluoroalkyl.

In an additional embodiment is the compound of Formula (G), wherein Wand Y are connected by a double or triple bond.

In an additional embodiment is the compound of Formula (G), wherein eachof R1, R2, R9 and R10 is hydrogen.

In an additional embodiment is the compound of Formula (G), wherein: pis 0, 1, 2 or 3; each R21 is independently alkyl or halo; R3, R4, R5 andR6 are each independently hydrogen, alkyl, halo or fluoroalkyl; and R15and R17 are each independently hydrogen, alkyl or halogen.

In an additional embodiment is the compound of Formula (A), wherein R11is alkyl. In an additional embodiment W is —C(R14)(R15)-. In a furtherembodiment, R1, R2, R3, R4, R5 and R6 are each independently hydrogen oralkyl.

In an additional embodiment, Z is —C(R16)(R17)- and the compound has thestructure of Formula (H):

as an isolated E or Z geometric isomer or a mixture of E and Z geometricisomers, as a tautomer or a mixture of tautomers, as a stereoisomer oras a pharmaceutically acceptable salt, hydrate, solvate, N-oxide orprodrug thereof, wherein:R1 and R2 are each the same or different and independently hydrogen oralkyl;R3, R4, R5 and R6 are each the same or different and independentlyhydrogen, halogen, nitro, —NH2, —NHR13, —N(R13)2, —OR12, alkyl orfluoroalkyl;R7 and R8 are each the same or different and independently hydrogen oralkyl; or R7 and R8 together with the carbon atom to which they areattached, form a carbocyclyl or heterocyclyl; or R7 and R8 together forman imino;R9 is hydrogen, alkyl, carbocyclyl, heterocyclyl, —C(═O)R13, —SO2R13,—CO2R13, —CONH2, —CON(R13)2 or —CON(H)R13;R10 is hydrogen or alkyl; or R9 and R10, together with the nitrogen atomto which they are attached, form an N-heterocyclyl;R11 is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;each R12 is independently selected from hydrogen or alkyl;each R13 is independently selected from alkyl, carbocyclyl,heterocyclyl, aryl or heteroaryl;R16 and R17 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19, carbocyclyl orheterocyclyl;each R18 and R19 is independently selected from hydrogen, alkyl,carbocyclyl, or —C(═O)R13, —SO2R13, —CO2R13, —CONH2, —CON(R13)2 or—CON(H)R13; or R18 and R19, together with the nitrogen atom to whichthey are attached, form an N-heterocyclyl.

In a further embodiment is the compound of Formula (H), wherein R1, R2,R3, R4, R5 and R6 are all hydrogen; R11 is aryl or carbocyclyl; and R16and R17 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl or —OR12.

In a further embodiment is the compound of Formula (A), wherein Z is abond and the compound has the structure of Formula (J):

as an isolated E or Z geometric isomer or a mixture of E and Z geometricisomers, as a tautomer or a mixture of tautomers, as a stereoisomer oras a pharmaceutically acceptable salt, hydrate, solvate, N-oxide orprodrug thereof, wherein:R1 and R2 are each the same or different and independently hydrogen oralkyl;R3, R4, R5 and R6 are each the same or different and independentlyhydrogen, halogen, nitro, —NH2, —NHR13, —N(R13)2, —OR12, alkyl orfluoroalkyl;R7 and R8 are each the same or different and independently hydrogen oralkyl; or R7 and R8 together with the carbon atom to which they areattached, form a carbocyclyl or heterocyclyl; or R7 and R8 together forman imino;R9 is hydrogen, alkyl, carbocyclyl, heterocyclyl, —C(═O)R13, —SO2R13,—CO2R13, —CONH2, —CON(R13)2 or —CON(H)R13;R10 is hydrogen or alkyl; or R9 and R10, together with the nitrogen atomto which they are attached, form an N-heterocyclyl;R11 is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;each R12 is independently selected from hydrogen or alkyl; andeach R13 is independently selected from alkyl, carbocyclyl,heterocyclyl, aryl or heteroaryl.

In a further embodiment is the compound of Formula (J), wherein

R1, R2, R3, R4, R5 and R6 are hydrogen; and

R11 is aryl or carbocyclyl.

In a further embodiment is the compound of Formula (A), wherein Z is—C(R14)(R15)-C(R16)(R17)-C(R21)(R22)- and the compound has the structureof Formula (K):

as an isolated E or Z geometric isomer or a mixture of E and Z geometricisomers, as a tautomer or a mixture of tautomers, as a stereoisomer oras a pharmaceutically acceptable salt, hydrate, solvate, N-oxide orprodrug thereof, wherein:R1 and R2 are each the same or different and independently hydrogen oralkyl;R3, R4, R5 and R6 are each the same or different and independentlyhydrogen, halogen, nitro, —NH2, —NHR13, —N(R13)2, —OR12, alkyl orfluoroalkyl;R7 and R8 are each the same or different and independently hydrogen oralkyl; or R7 and R8 together with the carbon atom to which they areattached, form a carbocyclyl or heterocyclyl; or R7 and R8 together forman imino;R9 is hydrogen, alkyl, carbocyclyl, heterocyclyl, —C(═O)R13, —SO2R13,—CO2R13, —CONH2, —CON(R13)2 or —CON(H)R13;R10 is hydrogen or alkyl; or R9 and R10, together with the nitrogen atomto which they are attached, form an N-heterocyclyl;R11 is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;each R12 is independently selected from hydrogen or alkyl;each R13 is independently selected from alkyl, carbocyclyl,heterocyclyl, aryl or heteroaryl;R14 and R15 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19, carbocyclyl orheterocyclyl; or R14 and R15 together form an oxo, an imino, an oximo,or a hydrazino;R16 and R17 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19, carbocyclyl orheterocyclyl; or R16 and R17 together form an oxo;each R18 and R19 is independently selected from hydrogen, alkyl,carbocyclyl, or —C(═O)R13, —SO2R13, —CO2R13, —CONH2, —CON(R13)2 or—CON(H)R13; or R18 and R19, together with the nitrogen atom to whichthey are attached, form an N-heterocyclyl; andR21 and R22 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19, carbocyclyl orheterocyclyl.In a further embodiment is the compound of Formula (K), whereinR1, R2, R3, R4, R5 and R6 are hydrogen;R11 is aryl or carbocyclyl;14 and R15 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl or —OR12; andR16, R17, R21 and R22 are each independently hydrogen or alkyl.

In a specific embodiment is the compound selected from:

-   (E)-3-(3-(2,6-dimethylstyryl)phenyl)propan-1-amine;-   (Z)-3-(3-(2,6-dimethylstyryl)phenyl)propan-1-amine;-   (E)-3-(3-(2-methylstyryl)phenyl)propan-1-amine;-   (Z)-3-(3-(2-methylstyryl)phenyl)propan-1-amine;-   (E)-3-(3-(2,6-dimethylstyryl)-2-methylphenyl)propan-1-amine;-   (Z)-3-(3-(2,6-dimethylstyryl)-2-methylphenyl)propan-1-amine;-   (E/Z)-3-(3-(2-ethyl-6-methylstyryl)phenyl)propan-1-amine;-   (E/Z)-3-(3-(2,5-dimethylstyryl)phenyl)propan-1-amine;-   (E/Z)-3-(3-(2,4-dimethylstyryl)phenyl)propan-1-amine;-   (E)-3-(3-(2,4,6-trimethylstyryl)phenyl)propan-1-amine;-   (E/Z)-3-(3-(2-ethylstyryl)phenyl)propan-1-amine;-   (E/Z)-3-(3-(2-ethynylstyryl)phenyl)propan-1-amine;-   (E/Z)-3-(3-(3,4-dimethylstyryl)phenyl)propan-1-amine;-   (E/Z)-3-(3-(2-isopropylstyryl)phenyl)propan-1-amine;-   (E/Z)-4-(3-(3,5-dimethylstyryl)phenyl)propan-1-amine;-   (E/Z)-4-(3-(2-methoxystyryl)phenyl)propan-1-amine;-   (E)-3-(3-(2,6-dichlorostyryl)phenyl)propan-1-amine;-   (E/Z)-3-(3-(2,3-dimethylstyryl)phenyl)propan-1-amine;-   (E)-3-(3-(2,6-dimethylstyryl)-4-fluorophenyl)propan-1-amine;-   (E/Z)-3-(3-(2-(trifluoromethyl)styryl)phenyl)propan-1-amine;-   (E)-3-(3-(2,6-dimethoxystyryl)phenyl)propan-1-amine;-   (E)-3-(3-(2,6-bis(trifluoromethyl) styryl)phenyl)propan-1-amine;-   (E)-3-amino-1-(3-(2,6-dichlorostyryl)phenyl)propan-1-ol;-   (E)-3-amino-1-(3-(2-chloro-6-methylstyryl)phenyl)propan-1-ol;-   (E)-2-(3-(3-aminopropyl)styryl)phenol;-   (E)-3-(5-(2,6-dichlorostyryl)-2-methoxyphenyl)propan-1-amine;-   (R,E)-1-amino-3-(3-(2,6-dichlorostyryl)phenyl)propan-2-ol;-   (S,E)-1-amino-3-(3-(2,6-dichlorostyryl)phenyl)propan-2-ol;-   (E/Z)-(3-(3-(2,6-diethoxystyryl)phenyl)propan-1-amine;-   (E)-3-(3-(2-ethoxystyryl)phenyl)propan-1-amine;-   (E/Z)-3-(3-(2-isopropoxystyryl)phenyl)propan-1-amine;-   (E)-3-amino-1-(3-(2,6-dichlorostyryl)phenyl)propan-1-one;-   (E)-1-amino-3-(3-(2,6-dichlorostyryl)phenyl)propan-2-one;-   (R,E)-3-amino-1-(3-(2,6-dichlorostyryl)phenyl)propan-1-ol;-   (S,E)-3-amino-1-(3-(2,6-dichlorostyryl)phenyl)propan-1-ol;-   (S,E)-3-(3-(2,6-dichlorostyryl)phenyl)-2-fluoropropan-1-amine;-   (E)-3-(3-(2,6-dichlorostyryl)phenyl)-2,2-difluoropropan-1-amine;-   (Z)-3-(3-(2-(2-methoxyethoxy)styryl)phenyl)-propan-1-amine;-   (E)-3-(3-(3-methoxystyryl)phenyl)propan-1-amine;-   (Z)-3-(3-(4-chlorostyryl)phenyl)propan-1-amine;-   (E)-3-(3-(2-(biphenyl-2-yl)vinyl)phenyl)propan-1-amine;-   (E)-3-(3-(3-chlorostyryl)phenyl)propan-1-amine;-   (E)-3-(3-(2-butoxystyryl)phenyl)propan-1-amine;-   (E)-3-(3-(4-methoxystyryl)phenyl)propan-1-amine;-   (Z)-3-(3-(2-Propoxystyryl)phenyl)propan-1-amine;-   (E)-3-(5-(2-Chloro-6-(methylthio)styryl)-2-methoxyphenyl)propan-1-amine;-   (E)-3-(3-(2-(1-methoxynaphthalen-2-yl)vinyl)phenyl)propan-1-amine;-   (Z)-3-(3-(2-(naphthalen-1-yl)vinyl)phenyl)propan-1-amine;-   (Z)-3-(3-(2-(3-methoxynaphthalen-2-yl)vinyl)phenyl)propan-1-amine;-   (E/Z)-3-(3-(2-(2-methoxynaphthalen-1-yl)vinyl)phenyl)propan-1-amine;-   (E)-2-amino-N-(3-(2,6-dimethylstyryl)phenyl)acetamide;-   (E)-2-(3-(2,6-dimethylstyryl)phenylthio)ethanamine;-   (E)-2-(3-(2,6-dimethylstyryl)phenylsulfinyl)ethanamine;-   (E)-2-(3-(2,6-dimethylstyryl)phenylsulfonyl)ethanamine;-   (E)-3-(3-(2,6-dimethylstyryl)phenyl)prop-2-en-1-amine;-   (E)-4-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)morpholine;-   (Z)-4-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)morpholine;-   (E)-1-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)pyrrolidine;-   (Z)-1-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)pyrrolidine;-   (E)-1-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)piperidine;-   (Z)-1-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)piperidine;-   (E)-3-(3-(2,6-dimethylstyryl)phenyl)-N-methylpropan-1-amine;-   (Z)-3-(3-(2,6-dimethylstyryl)phenyl)-N-methylpropan-1-amine;-   (E)-3-(3-(2,6-dimethylstyryl)phenyl)-N,N-dimethylpropan-1-amine;-   (Z)-3-(3-(2,6-dimethylstyryl)phenyl)-N,N-dimethylpropan-1-amine;-   (E/Z)—N-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)acetamide;-   (E)-1-(3-(3-aminopropyl)styryl)cyclohexanol;-   (E)-1-(3-(3-aminopropyl)styryl)cyclopentanol;-   (E)-1-(3-(3-aminopropyl)styryl)cycloheptanol;-   (E)-3-(3-(2-cyclohexylvinyl)phenyl)propan-1-amine;-   (E)-3-(3-(2-cyclopentylvinyl)phenyl)propan-1-amine;-   (E)-3-(3-(2-cycloheptylvinyl)phenyl)propan-1-amine;-   (E)-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine;-   (E)-3-amino-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-ol;-   (S,E)-3-amino-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-ol;-   (R,E)-3-Amino-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-ol;-   (E)-2-methyl-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine;-   (E)-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-1-amine;-   (E)-3-(2-methyl-5-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine;-   (E/Z)-4-amino-2-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-2-ol;-   (E)-3-fluoro-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine;-   (E)-4-amino-1,1,1-trifluoro-2-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-2-ol;-   (E)-3-amino-2,2-dimethyl-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-ol;-   (E)-3-amino-2-methyl-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-ol;-   (E)-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propane-1,3-diamine;-   (E)-4,4,4-trifluoro-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-1-amine;-   (E)-3-methoxy-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine;-   (E)-4-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-2-amine;-   (E)-1-amino-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-2-ol;-   (E)-2-(3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propylamino)ethanol;-   (E)-3-methoxy-N-methyl-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine;-   (E)-3-amino-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-one;-   (E)-3-amino-2,2-dimethyl-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-one;-   (E)-3-amino-2-methyl-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-one;-   (E)-3-amino-2-fluoro-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-one;-   (E)-2-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenoxy)ethanamine;-   (E)-2-amino-N-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)acetamide;-   (E)-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)prop-2-yn-1-amine;-   (E)-2-fluoro-3-(3-((E)-2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)prop-2-en-1-amine;-   (E)-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)prop-2-yn-1-amine;-   (E)-2-fluoro-3-(3-((E)-2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)prop-2-en-1-amine;-   (E)-3-(3-(pent-1-enyl)phenyl)propan-1-amine;-   (E)-3-(3-(hept-1-enyl)phenyl)propan-1-amine;-   (E)-3-(3-(non-4-en-5-yl)phenyl)propan-1-amine;-   (E)-4-(3-(3-aminopropyl)phenyl)-2-methylbut-3-en-2-ol;-   (E)-4-(3-(3-aminopropyl)styryl)heptan-4-ol;-   (E)-1-(3-(3-aminopropyl)phenyl)hex-1-en-3-ol;-   (E)-4-(3-(2-aminoethoxy)styryl)heptan-4-ol;-   (E)-1-(3-(3-aminopropyl)phenyl)-3-ethylpent-1-en-3-ol;-   (E)-3-(3-(3-aminopropyl)phenyl)prop-2-en-1-ol;-   (E)-3-(3-(3-methoxyprop-1-enyl)phenyl)propan-1-amine;-   (E)-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)methanamine;-   (E)-1-(3-(2,6-dimethylstyryl)phenyl)-N,N-dimethylmethanamine;-   (E)-4-(3-(2,6-dimethylstyryl)phenyl)butan-1-amine;-   (E)-2-(3-(2,6-dimethylstyryl)phenyl)ethanamine;-   (E)-2-(3-(2,6-dimethylstyryl)benzylamino)ethanol;-   (E)-3-(3-(2,6-dimethylstyryl)phenyl)-3-hydroxypropanimidamide;-   (E)-3-hydroxy-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propanimidamide;-   (Z)-3-amino-1-(3-(2,6-dimethylstyryl)phenyl)propan-1-one oxime;-   (Z)-3-amino-1-(3-((E)-2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-one    oxime;-   (Z)-3-(3-(2,6-dimethylstyryl)phenyl)-3-hydrazonopropan-1-amine;-   (Z)-3-hydrazono-3-(3-((E)-2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine;-   (E)-2-(3-(2,6-dimethylstyryl)phenyl)ethanamine;-   (E)-2-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)ethanamine;-   (E)-2-amino-1-(3-(2,6-dimethylstyryl)phenyl)ethanol;-   (E)-2-amino-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)ethanol;-   (E)-1-(3-(3-aminopropyl)phenyl)-3-methylhex-1-en-3-ol; and-   (E)-1-(3-(3-aminopropyl)phenyl)-3-ethylhex-1-en-3-ol.

In an additional embodiment is a pharmaceutical composition comprising apharmaceutically acceptable carrier and a compound disclosed herein,including compound of Formula (A)-(K).

In yet another embodiment is a compound that inhibits 11-cis-retinolproduction with an IC50 of about 1 uM or less when assayed in vitro,utilizing extract of cells that express RPE65 and LRAT, wherein theextract further comprises CRALBP, wherein the compound is stable insolution for at least about 1 week at room temperature. In a specificembodiment, the compound inhibits 11-cis-retinol production with an IC50of about 100 nM or less when assayed in vitro, utilizing extract ofcells that express RPE65 and LRAT, wherein the extract further comprisesCRALBP, wherein the compound is stable in solution for at least about 1week at room temperature. In a further embodiment, the compound inhibits11-cis-retinol production with an IC50 of about 10 nM or less whenassayed in vitro, utilizing extract of cells that express RPE65 andLRAT, wherein the extract further comprises CRALBP, wherein the compoundis stable in solution for at least about 1 week at room temperature.

In an additional embodiment is a non-retinoid compound that inhibits anisomerase reaction resulting in production of 11-cis retinol, whereinsaid isomerase reaction occurs in RPE, and wherein said compound has anED50 value of 1 mg/kg or less when administered to a subject. In afurther embodiment is a non-retinoid compound of wherein the ED50 valueis measured after administering a single dose of the compound to saidsubject for about 2 hours or longer. In an additional embodiment thecompound is a styrenyl compound. In a further embodiment the compound isa non-retinoid compound.

In a further embodiment is a pharmaceutical composition comprising apharmaceutically acceptable carrier and a compound that inhibits11-cis-retinol production with an IC50 of about 1 uM or less whenassayed in vitro, utilizing extract of cells that express RPE65 andLRAT, wherein the extract further comprises CRALBP, wherein the compoundis stable in solution for at least about 1 week at room temperature. Inan additional embodiment is a pharmaceutical composition comprising apharmaceutically acceptable carrier and a non-retinoid compound thatinhibits an isomerase reaction resulting in production of 11-cisretinol, wherein said isomerase reaction occurs in RPE, and wherein saidcompound has an ED50 value of 1 mg/kg or less when administered to asubject.

In another embodiment, the present invention provides a method ofmodulating chromophore flux in a retinoid cycle comprising introducinginto a subject a compound disclosed herein, including a compound of anyone of Formula (A)-(K). In a further embodiment the method results in areduction of lipofuscin pigment accumulated in an eye of the subject. Inyet another embodiment the lipofuscin pigment isN-retinylidene-N-retinyl-ethanolamine (A2E).

In yet another embodiment is a method for treating an ophthalmic diseaseor disorder in a subject, comprising administering to the subjectcompounds or the pharmaceutical composition described herein. In afurther embodiment, the ophthalmic disease or disorder is age-relatedmacular degeneration or Stargardt's macular dystrophy. In yet anotherembodiment the method results in a reduction of lipofuscin pigmentaccumulated in an eye of the subject. In yet another embodiment thelipofuscin pigment is N-retinylidene-N-retinyl-ethanolamine (A2E).

In additional embodiments, the ophthalmic disease or disorder isselected from retinal detachment, hemorrhagic retinopathy, retinitispigmentosa, cone-rod dystrophy, Sorsby's fundus dystrophy, opticneuropathy, inflammatory retinal disease, diabetic retinopathy, diabeticmaculopathy, retinal blood vessel occlusion, retinopathy of prematurity,or ischemia reperfusion related retinal injury, proliferativevitreoretinopathy, retinal dystrophy, hereditary optic neuropathy,Sorsby's fundus dystrophy, uveitis, a retinal injury, a retinal disorderassociated with Alzheimer's disease, a retinal disorder associated withmultiple sclerosis, a retinal disorder associated with Parkinson'sdisease, a retinal disorder associated with viral infection, a retinaldisorder related to light overexposure, myopia, and a retinal disorderassociated with AIDS.

In a further embodiment is a method of inhibiting dark adaptation of arod photoreceptor cell of the retina comprising contacting the retinawith a compound disclosed herein, including a compound of any one ofFormula (A)-(K).

In an additional embodiment is a method of inhibiting regeneration ofrhodopsin in a rod photoreceptor cell of the retina comprisingcontacting the retina with the compound of Formula (A), a compound thatinhibits 11-cis-retinol production with an IC50 of about 1 uM or lesswhen assayed in vitro, utilizing extract of cells that express RPE65 andLRAT, wherein the extract further comprises CRALBP, wherein the compoundis stable in solution for at least about 1 week at room temperature, ora A non-retinoid compound that inhibits an isomerase reaction resultingin production of 11-cis retinol, wherein said isomerase reaction occursin RPE, and wherein said compound has an ED50 value of 1 mg/kg or lesswhen administered to a subject.

In a further embodiment is a method of reducing ischemia in an eye of asubject comprising administering to the subject the pharmaceuticalcomposition of the compound of Formula (A), a compound that inhibits11-cis-retinol production with an IC50 of about 1 uM or less whenassayed in vitro, utilizing extract of cells that express RPE65 andLRAT, wherein the extract further comprises CRALBP, wherein the compoundis stable in solution for at least about 1 week at room temperature, ora A non-retinoid compound that inhibits an isomerase reaction resultingin production of 11-cis retinol, wherein said isomerase reaction occursin RPE, and wherein said compound has an ED50 value of 1 mg/kg or lesswhen administered to a subject. In a further embodiment, thepharmaceutical composition is administered under conditions and at atime sufficient to inhibit dark adaptation of a rod photoreceptor cell,thereby reducing ischemia in the eye.

In a further embodiment is a method of inhibiting neovascularization inthe retina of an eye of a subject comprising administering to thesubject the pharmaceutical composition of a compound of Formula (A). Ina specific embodiment, the pharmaceutical composition is administeredunder conditions and at a time sufficient to inhibit dark adaptation ofa rod photoreceptor cell, thereby inhibiting neovascularization in theretina.

In a further embodiment is a method of inhibiting degeneration of aretinal cell in a retina comprising contacting the retina with thecompound of the compound of Formula (A), a compound that inhibits11-cis-retinol production with an IC50 of about 1 uM or less whenassayed in vitro, utilizing extract of cells that express RPE65 andLRAT, wherein the extract further comprises CRALBP, wherein the compoundis stable in solution for at least about 1 week at room temperature, ora non-retinoid compound that inhibits an isomerase reaction resulting inproduction of 11-cis retinol, wherein said isomerase reaction occurs inRPE, and wherein said compound has an ED50 value of 1 mg/kg or less whenadministered to a subject. In a further embodiment, the pharmaceuticalcomposition is administered under conditions and at a time sufficient toinhibit dark adaptation of a rod photoreceptor cell, thereby reducingischemia in the eye. In a specific embodiment is the method wherein theretinal cell is a retinal neuronal cell. In a certain embodiment, theretinal neuronal cell is a photoreceptor cell.

Accordingly, in one embodiment, a compound is provided that has astructure of Formula (I):

as an isolated E or Z stereoisomer or a mixture of E and Zstereoisomers, as a tautomer or a mixture of tautomers, or as apharmaceutically acceptable salt, hydrate, solvate, N-oxide or prodrugthereof, wherein:

R₁ and R₂ are each the same or different and independently hydrogen oralkyl;

R₃, R₄, R₅ and R₆ are each the same or different and independentlyhydrogen, halogen, —OR₁₂, alkyl or fluoroalkyl;

R₇ and R₈ are each the same or different and independently hydrogen oralkyl;

R₉ is hydrogen, alkyl, carbocyclyl or —C(═O)R₁₃;

R₁₀ is hydrogen or alkyl; or

R₉ and R₁₀, together with the nitrogen atom to which they are attached,form an N-heterocyclyl;

R₁₁ is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;

R₁₂ is hydrogen or alkyl;

R₁₃ is alkyl, carbocyclyl or aryl;

W is —C(R₁₄)(R₁₅)—, —O—, —S—, —S(═O)—, —S(═O)₂— or —N(R₁₂)—;

Y is —C(R₁₆)(R₁₇)—;

R₁₄ and R₁₅ are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR₁₂, —NR₁₈R₁₉ or carbocyclyl; or

R₁₄ and R₁₅ form an oxo;

R₁₆ and R₁₇ are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR₁₂, —NR₁₈R₁₉ or carbocyclyl; or

R₁₆ and R₁₇ form an oxo; or

R₁₄ and R₁₆ together form a direct bond to provide a double bondconnecting W and Y; or

R₁₄ and R₁₆ together form a direct bond, and R₁₅ and R₁₇ together form adirect bond to provide a triple bond connecting W and Y;

R₁₈ and R₁₉ are each the same or different and independently hydrogen,alkyl, carbocyclyl, or —C(═O)R₁₃,

provided that when R₁₁ is phenyl, the compound of Formula (I) is not:

-   2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]acetamide;-   (2S,3R)-amino-3-hydroxy-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)-ethenyl]phenyl]-butanamide;-   L-glutamic acid,    1-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]ester;-   glycine, 3-hydroxy-5-[(1E)-2-(4-hydroxyphenyl)ethenyl]phenyl ester;-   (2S)-2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]propanamide;-   (2S)-2-amino-3-hydroxy-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]propanamide;-   (2S)-2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]-4-methyl-pentanamide;-   (2S)-2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]-3-methyl-butanamide;    or-   2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenylbutanamide.

Also provided are compounds having structures of Formulae (II), (IIa),(IIb), (III) and (IIIa):

wherein, t, p, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, R₉, R₁₀, R₁₄, R₁₅, R₁₆,R₁₇, R₂₀, R₂₁, W and Y are as defined above and herein.

Another embodiment provides a pharmaceutical composition comprising apharmaceutically acceptable carrier and a compound having a structure ofFormula (I):

as an isolated E or Z stereoisomer or a mixture of E and Zstereoisomers, as a tautomer or a mixture of tautomers, or as apharmaceutically acceptable salt, hydrate, solvate, N-oxide or prodrugthereof,

wherein, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, R₉, R₁₀, R₁₁, W and Y are asdefined herein.

Also provided are pharmaceutical compositions comprising a compoundhaving a structure of any of Formulae (II), (IIa), (IIb), (III) and(IIIa):

wherein, t, p, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, R₉, R₁₀, R₁₄, R₁₅, R₁₆,R₁₇, R₂₀, R₂₁, W and Y are as defined above and herein.

Also provided is a method for treating an ophthalmic disease or disorderin a subject comprising administering to the subject the pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier and acompound having a structure of any of Formulae (I), (II), (IIa), (IIb),(III) and (IIIa). In certain embodiments, the ophthalmic disease ordisorder is macular degeneration (i.e., age-related maculardegeneration) or Stargardt's disease. In other certain embodiments, theophthalmic disease or disorder is retinal detachment, hemorrhagicretinopathy, retinitis pigmentosa, optic neuropathy, inflammatoryretinal disease, proliferative vitreoretinopathy, retinal dystrophy,hereditary optic neuropathy, Sorsby's fundus dystrophy, uveitis, aretinal injury, a retinal disorder associated with Alzheimer's disease,a retinal disorder associated with multiple sclerosis, a retinaldisorder associated with Parkinson's disease, a retinal disorderassociated with viral infection, a retinal disorder related to lightoverexposure, and a retinal disorder associated with AIDS.

Also provided is a method of inhibiting at least one visual cycletrans-cis isomerase (also including trans-cis isomerohydrolase) in asubject comprising administering to the subject a pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier and acompound having a structure of any of Formulae (I), (II), (IIa), (IIb),(III) and (IIIa).

In certain particular embodiments of the aforementioned methods, thesubject is human. In other particular embodiments, accumulation oflipofuscin pigment is inhibited in an eye of the subject, wherein incertain embodiments the lipofuscin pigment isN-retinylidene-N-retinyl-ethanolamine (A2E). In another embodiment,degeneration of a retinal cell is inhibited. In particular embodiments,the retinal cell is a retinal neuronal cell, and other particularembodiments, the retinal neuronal cell is any one of an amacrine cell, aphotoreceptor cell, a horizontal cell, a ganglion cell, or a bipolarcell.

Also provided is a method of inhibiting at least one visual cycletrans-cis isomerase (also including trans-cis isomerohydrolase) in acell comprising contacting the cell with a compound having a structureof any of Formulae (I), (II), (IIa), (IIb), (III) and (IIIa), therebyinhibiting the at least one visual cycle trans-cis isomerase. In anotherembodiment, degeneration of a retinal cell is inhibited. In particularembodiments, the retinal cell is a retinal neuronal cell, and otherparticular embodiments, the retinal neuronal cell is any one of anamacrine cell, a photoreceptor cell, a horizontal cell, a ganglion cell,or a bipolar cell.

In another embodiment, a method is provided for treating an ophthalmicdisease or disorder in a subject, comprising administering to thesubject the pharmaceutical composition described above and hereincomprising at least one of the compounds having a structure of any ofFormulae (I), (II), (IIa), (IIb), (III) and (IIIa), wherein theophthalmic disease or disorder is selected from diabetic retinopathy,diabetic maculopathy, retinal blood vessel occlusion, retinopathy ofprematurit3y, or ischemia reperfusion related retinal injury.

INCORPORATION BY REFERENCE

All publications, patents, and patent applications mentioned in thisspecification are herein incorporated by reference to the same extent asif each individual publication, patent, or patent application wasspecifically and individually indicated to be incorporated by reference.

BRIEF DESCRIPTION OF THE DRAWINGS

The novel features of the invention are set forth with particularity inthe appended claims. A better understanding of the features andadvantages of the present invention will be obtained by reference to thefollowing detailed description that sets forth illustrative embodiments,in which the principles of the invention are utilized, and theaccompanying drawings of which:

FIG. 1 presents inhibition of isomerase activity by Compound A andCompound 1 over time in an in vivo mouse model. Five animals wereincluded in each treatment group. The error bars correspond to standarderror.

FIG. 2 shows concentration-dependent inhibition of isomerase activity byCompound A, Compound B, Compound 1 and Compound 25 in a mouse model.

DETAILED DESCRIPTION OF THE INVENTION

Styrenyl derivative compounds are described herein that inhibit anisomerization step of the retinoid cycle. These compounds andcompositions comprising these compounds may be useful for inhibitingdegeneration of retinal cells or for enhancing retinal cell survival.The compounds described herein may, therefore, be useful for treatingophthalmic diseases and disorders, such as age-related maculardegeneration and Stargardt's disease.

Styrenyl Derivative Compounds

In certain embodiments, styrenyl (i.e., vinyl benzene) derivativecompounds comprising a meta-substituted linkage terminating in anitrogen-containing moiety are provided. The nitrogen-containing moietycan be, for example, an amine, an amide or an N-heterocyclyl. Thelinking atoms form a combination of linearly constructed stable chemicalbonds, including single, double or triple carbon-carbon bonds,carbon-nitrogen bonds, nitrogen-nitrogen bonds, carbon-oxygen bonds,carbon-sulfur bonds, and the like.

In one embodiment is a compound having a structure of Formula (A):

as an isolated E or Z geometric isomer or a mixture of E and Z geometricisomers, as a tautomer or a mixture of tautomers, as a stereoisomer oras a pharmaceutically acceptable salt, hydrate, solvate, N-oxide orprodrug thereof, wherein: R1 and R2 are each the same or different andindependently hydrogen or alkyl; R3, R4, R5 and R6 are each the same ordifferent and independently hydrogen, halogen, nitro, —NH2, —NHR13,—N(R13)2, —OR12, alkyl or fluoroalkyl;R7 and R8 are each the same or different and independently hydrogen oralkyl; or R7 and R8 together with the carbon atom to which they areattached, form a carbocyclyl or heterocyclyl; or R7 and R8 together forman imino;R9 is hydrogen, alkyl, carbocyclyl, heterocyclyl, —C(═O)R13, —SO2R13,—CO2R13, —CONH2, —CON(R13)2 or —CON(H)R13;R10 is hydrogen or alkyl; or R9 and R10, together with the nitrogen atomto which they are attached, form an N-heterocyclyl;R11 is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;each R12 is independently selected from hydrogen or alkyl; each R13 isindependently selected from alkyl, carbocyclyl, heterocyclyl, aryl orheteroaryl;Z is a bond, Y or W—Y, wherein W is —C(R14)(R15)-, —O—, —S—, —S(═O)—,—S(═O)2- or —N(R12)-; Y is —C(R16)(R17)- or —C(R16)(R17)-C(R21)(R22)-;R14 and R15 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19, carbocyclyl orheterocyclyl; or R14 and R15 together form an oxo, an imino, an oximo,or a hydrazino; R16 and R17 are each the same or different andindependently hydrogen, halogen, alkyl, fluoroalkyl, —OR12, —NR18R19,carbocyclyl or heterocyclyl; or R16 and R17 together form an oxo; oroptionally, R14 and R16 together form a direct bond to provide a doublebond connecting W and Y; or optionally, R14 and R16 together form adirect bond, and R15 and R17 together form a direct bond to provide atriple bond connecting W and Y;each R18 and R19 is independently selected from hydrogen, alkyl,carbocyclyl, or —C(═O)R13, —SO2R13, —CO2R13, —CONH2, —CON(R13)2 or—CON(H)R13; or R18 and R19, together with the nitrogen atom to whichthey are attached, form an N-heterocyclyl; R21 and R22 are each the sameor different and independently hydrogen, halogen, alkyl, fluoroalkyl,—OR12, —NR18R19, carbocyclyl or heterocyclyl;provided that when R11 is phenyl, the compound of Formula (A) is not:

-   2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]acetamide;-   (2S,3R)-amino-3-hydroxy-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)-ethenyl]phenyl]-butanamide;-   L-glutamic acid,    1-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]ester;-   glycine, 3-hydroxy-5-[(1E)-2-(4-hydroxyphenyl)ethenyl]phenyl ester;-   (2S)-2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]propanamide;-   (2S)-2-amino-3-hydroxy-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]propanamide;-   (2S)-2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]-4-methyl-pentanamide;-   (2S)-2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]-3-methyl-butanamide;    or-   2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenylbutanamide.

In another embodiment is the compound wherein each of R1 and R8 ishydrogen, and the compound has a structure of Formula (B):

whereinR2 is H or C1-C6 alkyl;each of R3, R4, R5, and R6 is the same or different and independentlyhydrogen, halo, C1-C6 alkyl, or —OR12;R7 is H or C1-C6 alkyl;R9 is hydrogen, C1-C6 alkyl, —(CH2)nOH wherein n is 2-6, or —C(═O)R13;R10 is hydrogen or C1-C6 alkyl; or R9 and R10, together with thenitrogen atom to which they are attached, form an N-heterocyclyl;W is —C(R14)(R15)-, —O—, —S—, —S(═O)—, —S(═O)2- or —N(H)—;Y is —C(R16)(R17)-; each R12 is independently hydrogen or C1-C6 alkyl;each R13 is independently a C1-C6 alkyl; R14 and R15 are each the sameor different and independently selected from hydrogen, halogen, C1-C6alkyl, fluoroalkyl, —OR12, or —NH2; or R14 and R15 together form an oxo,an imino, an oximo, or a hydrazino; R16 and R17 are each the same ordifferent and independently hydrogen, halogen, C1-C6 alkyl, or —OR12; orR16 and R17 together form an oxo; or optionally, R14 and R16 togetherform a direct bond to provide a double bond connecting W and Y; oroptionally, R14 and R16 together form a direct bond, and R15 and R17together form a direct bond to provide a triple bond connecting W and Y;and R11 is selected from:a) alkyl;b) phenyl substituted with alkyl, —OR12, —O(CH2)mOCH3 wherein m is 1-6,alkenyl, alkynyl, halogen, fluoroalkyl, phenyl, —SCH3, or aralkyl;c) naphthenyl optionally substituted with alkyl, halogen, or —OR12;d) carbocyclyl; ore) cyclohexenyl optionally substituted with alkyl,provided that R11 is not 3,4,5-tri-methoxyphenyl.In a further embodiment is the compound of Formula (A) or (B) wherein R2is hydrogen or n-butyl. In an additional embodiment is the compound ofFormula (A) or (B) wherein halogen is fluoro or chloro.

In yet another embodiment is the compound of either Formula (A) or (B)wherein each of R3, R4, R5, and R6 is the same or different andindependently hydrogen, halogen, methyl, or methoxy.

In an additional embodiment is the compound of Formula (A) or (B),wherein at least two of R3, R4, R5, and R6 are hydrogen.

In a further embodiment is the compound of Formula (A) or (B), wherein Wis —C(R14)(R15)-, and wherein R14 and R15 are each the same or differentand independently hydrogen, fluoro, methyl, ethyl, trifluoromethyl, —OH,—OCH3, or —NH2.

In another embodiment is the compound of Formula (A) or (B), whereineach of R14 and R15 is hydrogen.

In an additional embodiment is the compound of Formula (A) or (B),wherein Y is —CH2-, —CH(CH3)-, —C(CH3)2-, —C(H)OH—, —C(H)F—, —CF2-, or—C(═O)—.

In yet another embodiment is the compound of Formula (A) or (B), whereinR11 is phenyl substituted with alkyl, —OR12, —O(CH2)nOCH3 wherein n is2-6, alkenyl, alkynyl, halogen, fluoroalkyl, phenyl, or —SCH3.

In one embodiment is the compound of Formula (A) or (B), wherein R11 isnaphthenyl substituted with —OR12, wherein R12 is hydrogen or C1-C6alkyl.

In an additional embodiment is the compound of Formula (A) or (B),wherein R11 is cyclohexenyl optionally substituted with C1-C6 alkyl. Insome embodiments R11 is tri-methyl-cyclohexenyl.

In additional embodiments is the compound of Formula (A) or (B), whereinR11 is alkyl optionally substituted with —OR12 wherein R12 is hydrogenor C1-C6 alkyl.

In an additional embodiment is the compound of Formula (A), wherein R11is aryl or carbocyclyl.

In an alternative embodiment is the compound of Formula (C), wherein R11is aryl:

as an isolated E or Z geometric isomer or a mixture of E and Z geometricisomers, as a tautomer or a mixture of tautomers, as a stereoisomer, oras a pharmaceutically acceptable salt, hydrate, solvate, N-oxide orprodrug thereof, wherein:R1 and R2 are each the same or different and independently hydrogen oralkyl;R3, R4, R5 and R6 are each the same or different and independentlyhydrogen, halogen, nitro, —NH2, —NHR13, —N(R13)2, —OR12, alkyl orfluoroalkyl;R7 and R8 are each the same or different and independently hydrogen oralkyl;R9 is hydrogen, alkyl, carbocyclyl or —C(═O)R13;R10 is hydrogen or alkyl; or R9 and R10, together with the nitrogen atomto which they are attached, form an N-heterocyclyl;each R12 is independently selected from hydrogen or alkyl;R13 is alkyl, carbocyclyl or aryl;W is —C(R14)(R15)-, —O—, —S—, —S(═O)—, —S(═O)2- or —N(R12)-;Y is —C(R16)(R17)-;R14 and R15 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19 or carbocyclyl; or R14 andR15 together form an oxo, an imino, an oximo, or a hydrazino;R16 and R17 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19 or carbocyclyl; or R16 andR17 together form an oxo;optionally, R14 and R16 together form a direct bond to provide a doublebond connecting W and Y; oroptionally, R14 and R16 together form a direct bond, and R15 and R17together form a direct bond to provide a triple bond connecting W and Y;each R18 and R19 is independently selected from hydrogen, alkyl,carbocyclyl, or —C(═O)R13;t is 0, 1, 2, 3, 4 or 5; andeach R20 is the same or different and independently selected from alkyl,—OR12, —SR12, alkenyl, alkynyl, halo, fluoroalkyl, aryl or aralkyl; ortwo adjacent R20 groups, together with the two carbon atoms to whichthey are attached, form a fused phenyl ring.

In an additional embodiment is the compound wherein each of R9 and R10is hydrogen and the compound has a structure of Formula (D):

In a further embodiment is the compound of Formula (E) wherein W is—C(R14)(R15)-:

as an isolated E or Z geometric isomer or a mixture of E and Z geometricisomers, as a tautomer or a mixture of tautomers, as a stereoisomer, oras a pharmaceutically acceptable salt, hydrate, solvate, N-oxide orprodrug thereof, wherein:R1 and R2 are each the same or different and independently hydrogen oralkyl;R3, R4, R5 and R6 are each the same or different and independentlyhydrogen, halogen, nitro, —NH2, —NHR13, —N(R13)2, —OR12, alkyl orfluoroalkyl;R7 and R8 are each the same or different and independently hydrogen oralkyl;each R12 is independently selected from hydrogen or alkyl;R13 is alkyl, carbocyclyl or aryl;R14 and R15 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19 or carbocyclyl; or R14 andR15 form an oxo, an imino, an oximo, or a hydrazino;R16 and R17 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19 or carbocyclyl; or R16 andR17 form an oxo;each R18 and R19 are each the same or different and independentlyhydrogen, alkyl, carbocyclyl, or —C(═O)R13;t is 0, 1, 2, 3, 4 or 5; andeach R20 is the same or different and independently alkyl, —OR12, —SR12,alkenyl, alkynyl, halo, fluoroalkyl, aryl or aralkyl, or two adjacentR20, together with the two carbon atoms to which they are attached, forma fused phenyl ring.In a additional embodiment is the compound of Formula (E) wherein t is0, 1, 2 or 3;each R20 is independently alkyl, —OR12, —SR12, alkynyl, phenyl, halo orfluoroalkyl; andR3, R4, R5 and R6 are each independently hydrogen, alkyl, —OR12, halo orfluoroalkyl.

In a further embodiment is the compound of Formula (E), wherein R7, R8,R14, R15, R16 and R17 are each independently hydrogen, halogen, alkyl or—OR12, wherein each R12 is independently selected from hydrogen oralkyl.

In an additional embodiment is the compound of Formula (E), wherein:

t is 2 or 3; two adjacent R20, together with the two carbon atoms towhich they are attached, form a fused phenyl ring; and

R3, R4, R5 and R6 are each independently hydrogen, alkyl, halo orfluoroalkyl.

In another embodiment is the compound of Formula (E), wherein R7, R8,R14, R15, R16 and R17 are each independently hydrogen, halogen, alkyl or—OR12.

In an additional embodiment is the compound of Formula (E), wherein W is—O—, —S—, —S(═O)—, —S(═O)2- or —N(R12)-.

In yet another embodiment is the compound of Formula (E), wherein:

t is 0, 1, 2 or 3;

each R20 is independently alkyl, —OR12 or halo; and

R3, R4, R5 and R6 are each independently hydrogen, alkyl or halo.

In a specific embodiment is the compound of Formula (E) wherein W and Yare connected by a double bond.

In an additional embodiment is the compound of Formula (E), wherein R9and R10 together with the nitrogen to which they are attached form aN-heterocyclyl.

In an additional embodiment is the compound of Formula (E), wherein theN-heterocyclyl is morpholinyl, pyrrolidinyl, piperidinyl or piperazinyl.

In an additional embodiment is the compound of Formula (E), wherein:

each of R1 and R2 is hydrogen;

t is 0, 1, 2 or 3;

each R20 is independently alkyl or halo; and

R3, R4, R5 and R6 are each independently hydrogen, alkyl or halo.

In another embodiment is the compound of Formula (E), wherein W is—C(R14)(R15)-.

In an additional embodiment is the compound of Formula (E), wherein R9is alkyl or —C(═O)R13, wherein R13 is alkyl, and R10 is hydrogen oralkyl.

In an additional embodiment is the compound of Formula (E), wherein:

each of R1 and R2 is hydrogen;

t is 0, 1, 2 or 3;

each R20 is independently alkyl or halo; and

R3, R4, R5 and R6 are each independently hydrogen, alkyl or halo.

In an additional embodiment is the compound of Formula (E), wherein W is—C(R14)(R15)-.

In an additional embodiment is the compound of Formula (A), wherein R11is 3-, 4-, 5-, 6-, 7- or 8-member cycloalkyl. In some embodiments R11 iscyclohexyl.

In an additional embodiment is the compound of Formula (A), wherein R11is 5-, 6-, or 7-member cycloalkenyl.

In an additional embodiment is the compound of Formula (F), wherein R11is cyclohexenyl:

as an isolated E or Z geometric isomer or a mixture of E and Z geometricisomers, as a tautomer or a mixture of tautomers, as a stereoisomer, oras a pharmaceutically acceptable salt, hydrate, solvate, N-oxide orprodrug thereof, wherein:R1 and R2 are each the same or different and independently hydrogen oralkyl;R3, R4, R5 and R6 are each the same or different and independentlyhydrogen, halogen, nitro, —NH2, —NHR13, —N(R13)2, —OR12, alkyl orfluoroalkyl;R7 and R8 are each the same or different and independently hydrogen oralkyl;R9 is hydrogen, alkyl, carbocyclyl or —C(═O)R13;R10 is hydrogen or alkyl; or R9 and R10, together with the nitrogen atomto which they are attached, form an N-heterocyclyl;each R12 is independently selected from hydrogen or alkyl;each R13 is independently selected from alkyl, carbocyclyl or aryl;W is —C(R14)(R15)-, —O—, —S—, —S(═O)—, —S(═O)2- or —N(R12)-;Y is —C(R16)(R17)-;R14 and R15 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19 or carbocyclyl; or R14 andR15 together form an oxo, an imino, an oximo, or a hydrazino;R16 and R17 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19 or carbocyclyl; or R16 andR17 together form an oxo;optionally, R14 and R16 together form a direct bond to provide a doublebond connecting W and Y; or R14 and R16 together form a direct bond, andR15 and R17 together form a direct bond to provide a triple bondconnecting W and Y;each R18 and R19 is independently selected from hydrogen, alkyl,carbocyclyl, or —C(═O)R13;p is 0, 1, 2, 3, 4, 5, 7, 8 or 9; andeach R21 is the same or different and independently selected from alkyl,—OR12, alkenyl, alkynyl, halo, fluoroalkyl or aralkyl.

In an additional embodiment is the compound wherein W is —C(R14)(R15)-,and the compound has a structure of Formula (G):

as an isolated E or Z geometric isomer or a mixture of E and Z geometricisomers, as a tautomer or a mixture of tautomers, as a stereoisomer, oras a pharmaceutically acceptable salt, hydrate, solvate, N-oxide orprodrug thereof, wherein:R1 and R2 are each the same or different and independently hydrogen oralkyl;R3, R4, R5 and R6 are each the same or different and independentlyhydrogen, halogen, nitro, —NH2, —NHR13, —N(R13)2, —OR12, alkyl orfluoroalkyl;R7 and R8 are each the same or different and independently hydrogen oralkyl;R9 is hydrogen, alkyl, carbocyclyl or —C(═O)R13;R10 is hydrogen or alkyl; or R9 and R10, together with the nitrogen atomto which they are attached, form an N-heterocyclyl;each R12 is independently selected from hydrogen or alkyl;each R13 is independently alkyl, carbocyclyl or aryl;R14 and R15 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19 or carbocyclyl; or R14 andR15 together form an oxo, an imino, an oximo, or a hydrazino;R16 and R17 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19 or carbocyclyl; or R16 andR17 together form an oxo;optionally, R14 and R16 together form a direct bond to provide a doublebond connecting W and Y; or R14 and R16 together form a direct bond, andR15 and R17 together form a direct bond to provide a triple bondconnecting W and Y;each R18 and R19 is independently selected from hydrogen, alkyl,carbocyclyl, or —C(═O)R13;p is 0, 1, 2, 3, 4, 5, 7, 8 or 9; andeach R21 is the same or different and independently alkyl, —OR12,alkenyl, alkynyl, halo, fluoroalkyl or aralkyl.

In an additional embodiment is the compound of Formula (G), wherein eachof R9 and R10 is hydrogen.

In an additional embodiment is the compound of Formula (G), wherein:

each of R1 and R2 is hydrogen;

p is 0, 1, 2 or 3;

each R21 is independently alkyl, halo or fluoroalkyl; and

each of R3, R4, R5 and R6 is independently hydrogen, alkyl, halo,fluoroalkyl or —OR12.

In an additional embodiment is the compound of Formula (G), wherein:

R7, R8, R14, R15, R16 and R17 are each independently hydrogen, halogen,alkyl, fluoroalkyl, —OR12 or —NR18R19, wherein each R12 is independentlyhydrogen or alkyl; and each R18 and R19 are independently hydrogen oralkyl.

In an additional embodiment is the compound of Formula (G), wherein R9is alkyl and R10 is hydrogen.

In an additional embodiment is the compound of Formula (G), wherein:

each of R1 and R2 is hydrogen;

p is 0, 1, 2 or 3;

each R21 is independently alkyl, halo or fluoroalkyl; and

each of R3, R4, R5 and R6 is independently hydrogen, alkyl, halo,fluoroalkyl or —OR12.

In an additional embodiment is the compound of Formula (G), wherein R7,R8, R14, R15, R16 and R17 are each independently hydrogen, halogen,alkyl, fluoroalkyl or —OR12, wherein each R12 is independently hydrogenor alkyl.

In an additional embodiment is the compound of Formula (G), wherein:

R7, R8, R16 and R17 are each independently hydrogen, halogen, alkyl,fluoroalkyl or —OR12, wherein R12 is hydrogen or alkyl; and

R14 and R15 together form oxo.

In an additional embodiment is the compound of Formula (G), wherein W is—NH— or —O—.

In an additional embodiment is the compound of Formula (G), wherein eachof R1, R2, R9 and R10 is hydrogen.

In an additional embodiment is the compound of Formula (G), wherein:

p is 0, 1, 2 or 3;

each R21 is independently alkyl or halo; and

R3, R4, R5 and R6 are each independently hydrogen, alkyl, halo orfluoroalkyl.

In an additional embodiment is the compound of Formula (G), wherein Wand Y are connected by a double or triple bond.

In an additional embodiment is the compound of Formula (G), wherein eachof R1, R2, R9 and R10 is hydrogen.

In an additional embodiment is the compound of Formula (G), wherein:

p is 0, 1, 2 or 3;

each R21 is independently alkyl or halo;

R3, R4, R5 and R6 are each independently hydrogen, alkyl, halo orfluoroalkyl; and

R15 and R17 are each independently hydrogen, alkyl or halogen.

In an additional embodiment is the compound of Formula (A), wherein R11is alkyl. In an additional embodiment W is —C(R14)(R15)-. In a furtherembodiment, R1, R2, R3, R4, R5 and R6 are each independently hydrogen oralkyl.

In an additional embodiment, Z is —C(R16)(R17)- and the compound has thestructure of Formula (H):

as an isolated E or Z geometric isomer or a mixture of E and Z geometricisomers, as a tautomer or a mixture of tautomers, as a stereoisomer oras a pharmaceutically acceptable salt, hydrate, solvate, N-oxide orprodrug thereof, wherein:R1 and R2 are each the same or different and independently hydrogen oralkyl;R3, R4, R5 and R6 are each the same or different and independentlyhydrogen, halogen, nitro, —NH2, —NHR13, —N(R13)2, —OR12, alkyl orfluoroalkyl;R7 and R8 are each the same or different and independently hydrogen oralkyl; or R7 and R8 together with the carbon atom to which they areattached, form a carbocyclyl or heterocyclyl; or R7 and R8 together forman imino;R9 is hydrogen, alkyl, carbocyclyl, heterocyclyl, —C(═O)R13, —SO2R13,—CO2R13, —CONH2, —CON(R13)2 or —CON(H)R13;R10 is hydrogen or alkyl; or R9 and R10, together with the nitrogen atomto which they are attached, form an N-heterocyclyl;R11 is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;each R12 is independently selected from hydrogen or alkyl;each R13 is independently selected from alkyl, carbocyclyl,heterocyclyl, aryl or heteroaryl;R16 and R17 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19, carbocyclyl orheterocyclyl;each R18 and R19 is independently selected from hydrogen, alkyl,carbocyclyl, or —C(═O)R13, —SO2R13, —CO2R13, —CONH2, —CON(R13)2 or—CON(H)R13; or R18 and R19, together with the nitrogen atom to whichthey are attached, form an N-heterocyclyl.

In a further embodiment is the compound of Formula (H), wherein

R1, R2, R3, R4, R5 and R6 are all hydrogen;

R11 is aryl or carbocyclyl; and

R16 and R17 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl or —OR12.

In a further embodiment is the compound of Formula (A), wherein Z is abond and the compound has the structure of Formula (J):

as an isolated E or Z geometric isomer or a mixture of E and Z geometricisomers, as a tautomer or a mixture of tautomers, as a stereoisomer oras a pharmaceutically acceptable salt, hydrate, solvate, N-oxide orprodrug thereof, wherein:R1 and R2 are each the same or different and independently hydrogen oralkyl;R3, R4, R5 and R6 are each the same or different and independentlyhydrogen, halogen, nitro, —NH2, —NHR13, —N(R13)2, —OR12, alkyl orfluoroalkyl;R7 and R8 are each the same or different and independently hydrogen oralkyl; or R7 and R8 together with the carbon atom to which they areattached, form a carbocyclyl or heterocyclyl; or R7 and R8 together forman imino;R9 is hydrogen, alkyl, carbocyclyl, heterocyclyl, —C(═O)R13, —SO2R13,—CO2R13, —CONH2, —CON(R13)2 or —CON(H)R13;R10 is hydrogen or alkyl; or R9 and R10, together with the nitrogen atomto which they are attached, form an N-heterocyclyl;R11 is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;each R12 is independently selected from hydrogen or alkyl; andeach R13 is independently selected from alkyl, carbocyclyl,heterocyclyl, aryl or heteroaryl.

In a further embodiment is the compound of Formula (J), wherein

R1, R2, R3, R4, R5 and R6 are hydrogen; and

R11 is aryl or carbocyclyl.

In a further embodiment is the compound of Formula (A), wherein Z is—C(R14)(R15)-C(R16)(R17)-C(R21)(R22)- and the compound has the structureof Formula (K):

as an isolated E or Z geometric isomer or a mixture of E and Z geometricisomers, as a tautomer or a mixture of tautomers, as a stereoisomer oras a pharmaceutically acceptable salt, hydrate, solvate, N-oxide orprodrug thereof, wherein:R1 and R2 are each the same or different and independently hydrogen oralkyl;R3, R4, R5 and R6 are each the same or different and independentlyhydrogen, halogen, nitro, —NH2, —NHR13, —N(R13)2, —OR12, alkyl orfluoroalkyl;R7 and R8 are each the same or different and independently hydrogen oralkyl; or R7 and R8 together with the carbon atom to which they areattached, form a carbocyclyl or heterocyclyl; or R7 and R8 together forman imino;R9 is hydrogen, alkyl, carbocyclyl, heterocyclyl, —C(═O)R13, —SO2R13,—CO2R13, —CONH2, —CON(R13)2 or —CON(H)R13;R10 is hydrogen or alkyl; or R9 and R10, together with the nitrogen atomto which they are attached, form an N-heterocyclyl;R11 is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;each R12 is independently selected from hydrogen or alkyl;each R13 is independently selected from alkyl, carbocyclyl,heterocyclyl, aryl or heteroaryl;R14 and R15 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19, carbocyclyl orheterocyclyl; or R14 and R15 together form an oxo, an imino, an oximo,or a hydrazino;R16 and R17 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19, carbocyclyl orheterocyclyl; or R16 and R17 together form an oxo;each R18 and R19 is independently selected from hydrogen, alkyl,carbocyclyl, or —C(═O)R13, —SO2R13, —CO2R13, —CONH2, —CON(R13)2 or—CON(H)R13; or R18 and R19, together with the nitrogen atom to whichthey are attached, form an N-heterocyclyl; andR21 and R22 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR12, —NR18R19, carbocyclyl orheterocyclyl.In a further embodiment is the compound of Formula (K), whereinR1, R2, R3, R4, R5 and R6 are hydrogen;R11 is aryl or carbocyclyl;14 and R15 are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl or —OR12; andR16, R17, R21 and R22 are each independently hydrogen or alkyl.

In a specific embodiment is the compound selected from:

-   (E)-3-(3-(2,6-dimethylstyryl)phenyl)propan-1-amine;-   (Z)-3-(3-(2,6-dimethylstyryl)phenyl)propan-1-amine;-   (E)-3-(3-(2-methylstyryl)phenyl)propan-1-amine;-   (Z)-3-(3-(2-methylstyryl)phenyl)propan-1-amine;-   (E)-3-(3-(2,6-dimethylstyryl)-2-methylphenyl)propan-1-amine;-   (Z)-3-(3-(2,6-dimethylstyryl)-2-methylphenyl)propan-1-amine;-   (E/Z)-3-(3-(2-ethyl-6-methylstyryl)phenyl)propan-1-amine;-   (E/Z)-3-(3-(2,5-dimethylstyryl)phenyl)propan-1-amine;-   (E/Z)-3-(3-(2,4-dimethylstyryl)phenyl)propan-1-amine;-   (E)-3-(3-(2,4,6-trimethylstyryl)phenyl)propan-1-amine;-   (E/Z)-3-(3-(2-ethylstyryl)phenyl)propan-1-amine;-   (E/Z)-3-(3-(2-ethynylstyryl)phenyl)propan-1-amine;-   (E/Z)-3-(3-(3,4-dimethylstyryl)phenyl)propan-1-amine;-   (E/Z)-3-(3-(2-isopropylstyryl)phenyl)propan-1-amine;-   (E/Z)-4-(3-(3,5-dimethylstyryl)phenyl)propan-1-amine;-   (E/Z)-4-(3-(2-methoxystyryl)phenyl)propan-1-amine;-   (E)-3-(3-(2,6-dichlorostyryl)phenyl)propan-1-amine;-   (E/Z)-3-(3-(2,3-dimethylstyryl)phenyl)propan-1-amine;-   (E)-3-(3-(2,6-dimethylstyryl)-4-fluorophenyl)propan-1-amine;-   (E/Z)-3-(3-(2-(trifluoromethyl)styryl)phenyl)propan-1-amine;-   (E)-3-(3-(2,6-dimethoxystyryl)phenyl)propan-1-amine;-   (E)-3-(3-(2,6-bis(trifluoromethyl) styryl)phenyl)propan-1-amine;-   (E)-3-amino-1-(3-(2,6-dichlorostyryl)phenyl)propan-1-ol;-   (E)-3-amino-1-(3-(2-chloro-6-methylstyryl)phenyl)propan-1-ol;-   (E)-2-(3-(3-aminopropyl)styryl)phenol;-   (E)-3-(5-(2,6-dichlorostyryl)-2-methoxyphenyl)propan-1-amine;-   (R,E)-1-amino-3-(3-(2,6-dichlorostyryl)phenyl)propan-2-ol;-   (S,E)-1-amino-3-(3-(2,6-dichlorostyryl)phenyl)propan-2-ol;-   (E/Z)-(3-(3-(2,6-diethoxystyryl)phenyl)propan-1-amine;-   (E)-3-(3-(2-ethoxystyryl)phenyl)propan-1-amine;-   (E/Z)-3-(3-(2-isopropoxystyryl)phenyl)propan-1-amine;-   (E)-3-amino-1-(3-(2,6-dichlorostyryl)phenyl)propan-1-one;-   (E)-1-amino-3-(3-(2,6-dichlorostyryl)phenyl)propan-2-one;-   (R,E)-3-amino-1-(3-(2,6-dichlorostyryl)phenyl)propan-1-ol;-   (S,E)-3-amino-1-(3-(2,6-dichlorostyryl)phenyl)propan-1-ol;-   (S,E)-3-(3-(2,6-dichlorostyryl)phenyl)-2-fluoropropan-1-amine;-   (E)-3-(3-(2,6-dichlorostyryl)phenyl)-2,2-difluoropropan-1-amine;-   (Z)-3-(3-(2-(2-methoxyethoxy)styryl)phenyl)-propan-1-amine;-   (E)-3-(3-(3-methoxystyryl)phenyl)propan-1-amine;-   (Z)-3-(3-(4-chlorostyryl)phenyl)propan-1-amine;-   (E)-3-(3-(2-(biphenyl-2-yl)vinyl)phenyl)propan-1-amine;-   (E)-3-(3-(3-chlorostyryl)phenyl)propan-1-amine;-   (E)-3-(3-(2-butoxystyryl)phenyl)propan-1-amine;-   (E)-3-(3-(4-methoxystyryl)phenyl)propan-1-amine;-   (Z)-3-(3-(2-Propoxystyryl)phenyl)propan-1-amine;-   (E)-3-(5-(2-Chloro-6-(methylthio)styryl)-2-methoxyphenyl)propan-1-amine;-   (E)-3-(3-(2-(1-methoxynaphthalen-2-yl)vinyl)phenyl)propan-1-amine;-   (Z)-3-(3-(2-(naphthalen-1-yl)vinyl)phenyl)propan-1-amine;-   (Z)-3-(3-(2-(3-methoxynaphthalen-2-yl)vinyl)phenyl)propan-1-amine;-   (E/Z)-3-(3-(2-(2-methoxynaphthalen-1-yl)vinyl)phenyl)propan-1-amine;-   (E)-2-amino-N-(3-(2,6-dimethylstyryl)phenyl)acetamide;-   (E)-2-(3-(2,6-dimethylstyryl)phenylthio)ethanamine;-   (E)-2-(3-(2,6-dimethylstyryl)phenylsulfinyl)ethanamine;-   (E)-2-(3-(2,6-dimethylstyryl)phenylsulfonyl)ethanamine;-   (E)-3-(3-(2,6-dimethylstyryl)phenyl)prop-2-en-1-amine;-   (E)-4-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)morpholine;-   (Z)-4-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)morpholine;-   (E)-1-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)pyrrolidine;-   (Z)-1-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)pyrrolidine;-   (E)-1-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)piperidine;-   (Z)-1-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)piperidine;-   (E)-3-(3-(2,6-dimethylstyryl)phenyl)-N-methylpropan-1-amine;-   (Z)-3-(3-(2,6-dimethylstyryl)phenyl)-N-methylpropan-1-amine;-   (E)-3-(3-(2,6-dimethylstyryl)phenyl)-N,N-dimethylpropan-1-amine;-   (Z)-3-(3-(2,6-dimethylstyryl)phenyl)-N,N-dimethylpropan-1-amine;-   (E/Z)—N-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)acetamide;-   (E)-1-(3-(3-aminopropyl)styryl)cyclohexanol;-   (E)-1-(3-(3-aminopropyl)styryl)cyclopentanol;-   (E)-1-(3-(3-aminopropyl)styryl)cycloheptanol;-   (E)-3-(3-(2-cyclohexylvinyl)phenyl)propan-1-amine;-   (E)-3-(3-(2-cyclopentylvinyl)phenyl)propan-1-amine;-   (E)-3-(3-(2-cycloheptylvinyl)phenyl)propan-1-amine;-   (E)-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine;-   (E)-3-amino-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-ol;-   (S,E)-3-amino-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-ol;-   (R,E)-3-Amino-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-ol;-   (E)-2-methyl-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine;-   (E)-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-1-amine;-   (E)-3-(2-methyl-5-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine;-   (E/Z)-4-amino-2-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-2-ol;-   (E)-3-fluoro-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine;-   (E)-4-amino-1,1,1-trifluoro-2-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-2-ol;-   (E)-3-amino-2,2-dimethyl-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-ol;-   (E)-3-amino-2-methyl-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-ol;-   (E)-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propane-1,3-diamine;-   (E)-4,4,4-trifluoro-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-1-amine;-   (E)-3-methoxy-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine;-   (E)-4-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-2-amine;-   (E)-1-amino-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-2-ol;-   (E)-2-(3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propylamino)ethanol;-   (E)-3-methoxy-N-methyl-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine;-   (E)-3-amino-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-one;-   (E)-3-amino-2,2-dimethyl-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-one;-   (E)-3-amino-2-methyl-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-one;-   (E)-3-amino-2-fluoro-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-one;-   (E)-2-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenoxy)ethanamine;-   (E)-2-amino-N-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)acetamide;-   (E)-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)prop-2-yn-1-amine;-   (E)-2-fluoro-3-(3-((E)-2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)prop-2-en-1-amine;-   (E)-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)prop-2-yn-1-amine;-   (E)-2-fluoro-3-(3-((E)-2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)prop-2-en-1-amine;-   (E)-3-(3-(pent-1-enyl)phenyl)propan-1-amine;-   (E)-3-(3-(hept-1-enyl)phenyl)propan-1-amine;-   (E)-3-(3-(non-4-en-5-yl)phenyl)propan-1-amine;-   (E)-4-(3-(3-aminopropyl)phenyl)-2-methylbut-3-en-2-ol;-   (E)-4-(3-(3-aminopropyl)styryl)heptan-4-ol;-   (E)-1-(3-(3-aminopropyl)phenyl)hex-1-en-3-ol;-   (E)-4-(3-(2-aminoethoxy)styryl)heptan-4-ol;-   (E)-1-(3-(3-aminopropyl)phenyl)-3-ethylpent-1-en-3-ol;-   (E)-3-(3-(3-aminopropyl)phenyl)prop-2-en-1-ol;-   (E)-3-(3-(3-methoxyprop-1-enyl)phenyl)propan-1-amine;-   (E)-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)methanamine;-   (E)-1-(3-(2,6-dimethylstyryl)phenyl)-N,N-dimethylmethanamine;-   (E)-4-(3-(2,6-dimethylstyryl)phenyl)butan-1-amine;-   (E)-2-(3-(2,6-dimethylstyryl)phenyl)ethanamine;-   (E)-2-(3-(2,6-dimethylstyryl)benzylamino)ethanol;-   (E)-3-(3-(2,6-dimethylstyryl)phenyl)-3-hydroxypropanimidamide;-   (E)-3-hydroxy-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propanimidamide;-   (Z)-3-amino-1-(3-(2,6-dimethylstyryl)phenyl)propan-1-one oxime;-   (Z)-3-amino-1-(3-((E)-2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-one    oxime;-   (Z)-3-(3-(2,6-dimethylstyryl)phenyl)-3-hydrazonopropan-1-amine;-   (Z)-3-hydrazono-3-(3-((E)-2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine;-   (E)-2-(3-(2,6-dimethylstyryl)phenyl)ethanamine;-   (E)-2-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)ethanamine;-   (E)-2-amino-1-(3-(2,6-dimethylstyryl)phenyl)ethanol;-   (E)-2-amino-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)ethanol;-   (E)-1-(3-(3-aminopropyl)phenyl)-3-methylhex-1-en-3-ol; and-   (E)-1-(3-(3-aminopropyl)phenyl)-3-ethylhex-1-en-3-ol.

In an additional embodiment is a pharmaceutical composition comprising apharmaceutically acceptable carrier and a compound disclosed herein,including a compound of any one of Formula (A)-(K).

In yet another embodiment is a compound that inhibits 11-cis-retinolproduction with an IC50 of about 1 uM or less when assayed in vitro,utilizing extract of cells that express RPE65 and LRAT, wherein theextract further comprises CRALBP, wherein the compound is stable insolution for at least about 1 week at room temperature. In a specificembodiment, the compound inhibits 11-cis-retinol production with an IC50of about 100 nM or less when assayed in vitro, utilizing extract ofcells that express RPE65 and LRAT, wherein the extract further comprisesCRALBP, wherein the compound is stable in solution for at least about 1week at room temperature. In a further embodiment, the compound inhibits11-cis-retinol production with an IC50 of about 10 nM or less whenassayed in vitro, utilizing extract of cells that express RPE65 andLRAT, wherein the extract further comprises CRALBP, wherein the compoundis stable in solution for at least about 1 week at room temperature.

In an additional embodiment is a non-retinoid compound that inhibits anisomerase reaction resulting in production of 11-cis retinol, whereinsaid isomerase reaction occurs in RPE, and wherein said compound has anED50 value of 1 mg/kg or less when administered to a subject. In afurther embodiment is a non-retinoid compound of wherein the ED50 valueis measured after administering a single dose of the compound to saidsubject for about 2 hours or longer. In an additional embodiment thecompound is a styrenyl compound. In a further embodiment the compound isa non-retinoid compound.

In a further embodiment is a pharmaceutical composition comprising apharmaceutically acceptable carrier and a compound that inhibits11-cis-retinol production with an IC50 of about 1 uM or less whenassayed in vitro, utilizing extract of cells that express RPE65 andLRAT, wherein the extract further comprises CRALBP, wherein the compoundis stable in solution for at least about 1 week at room temperature. Inan additional embodiment is a pharmaceutical composition comprising apharmaceutically acceptable carrier and a non-retinoid compound thatinhibits an isomerase reaction resulting in production of 11-cisretinol, wherein said isomerase reaction occurs in RPE, and wherein saidcompound has an ED50 value of 1 mg/kg or less when administered to asubject.

In a specific embodiment is a method of modulating chromophore flux in aretinoid cycle comprising introducing into a subject a compounddisclosed herein, including a compound of any one of Formula (A)-(K). Ina further embodiment the method results in a reduction of lipofuscinpigment accumulated in an eye of the subject. In yet another embodimentthe lipofuscin pigment is N-retinylidene-N-retinyl-ethanolamine (A2E).

In yet another embodiment is a method for treating an ophthalmic diseaseor disorder in a subject, comprising administering to the subject thepharmaceutical composition described herein. In a further embodiment,the ophthalmic disease or disorder is age-related macular degenerationor Stargardt's macular dystrophy. In yet another embodiment the methodresults in a reduction of lipofuscin pigment accumulated in an eye ofthe subject. In yet another embodiment the lipofuscin pigment isN-retinylidene-N-retinyl-ethanolamine (A2E).

In additional embodiments, the ophthalmic disease or disorder isselected from retinal detachment, hemorrhagic retinopathy, retinitispigmentosa, cone-rod dystrophy, Sorsby's fundus dystrophy, opticneuropathy, inflammatory retinal disease, diabetic retinopathy, diabeticmaculopathy, retinal blood vessel occlusion, retinopathy of prematurity,or ischemia reperfusion related retinal injury, proliferativevitreoretinopathy, retinal dystrophy, hereditary optic neuropathy,Sorsby's fundus dystrophy, uveitis, a retinal injury, a retinal disorderassociated with Alzheimer's disease, a retinal disorder associated withmultiple sclerosis, a retinal disorder associated with Parkinson'sdisease, a retinal disorder associated with viral infection, a retinaldisorder related to light overexposure, myopia, and a retinal disorderassociated with AIDS.

In a further embodiment is a method of inhibiting dark adaptation of arod photoreceptor cell of the retina comprising contacting the retinawith a compound disclosed herein, including a compound of any one ofFormula (A)-(K).

In an additional embodiment is a method of inhibiting regeneration ofrhodopsin in a rod photoreceptor cell of the retina comprisingcontacting the retina with the compound of Formula (A), a compound thatinhibits 11-cis-retinol production with an IC50 of about 1 uM or lesswhen assayed in vitro, utilizing extract of cells that express RPE65 andLRAT, wherein the extract further comprises CRALBP, wherein the compoundis stable in solution for at least about 1 week at room temperature, ora A non-retinoid compound that inhibits an isomerase reaction resultingin production of 11-cis retinol, wherein said isomerase reaction occursin RPE, and wherein said compound has an ED50 value of 1 mg/kg or lesswhen administered to a subject.

In a further embodiment is a method of reducing ischemia in an eye of asubject comprising administering to the subject the pharmaceuticalcomposition of the compound of Formula (A), a compound that inhibits11-cis-retinol production with an IC50 of about 1 uM or less whenassayed in vitro, utilizing extract of cells that express RPE65 andLRAT, wherein the extract further comprises CRALBP, wherein the compoundis stable in solution for at least about 1 week at room temperature, ora A non-retinoid compound that inhibits an isomerase reaction resultingin production of 11-cis retinol, wherein said isomerase reaction occursin RPE, and wherein said compound has an ED50 value of 1 mg/kg or lesswhen administered to a subject. In a further embodiment, thepharmaceutical composition is administered under conditions and at atime sufficient to inhibit dark adaptation of a rod photoreceptor cell,thereby reducing ischemia in the eye.

In a further embodiment is a method of inhibiting neovascularization inthe retina of an eye of a subject comprising administering to thesubject the pharmaceutical composition of a compound of Formula (A). Ina specific embodiment, the pharmaceutical composition is administeredunder conditions and at a time sufficient to inhibit dark adaptation ofa rod photoreceptor cell, thereby inhibiting neovascularization in theretina.

In a further embodiment is a method of inhibiting degeneration of aretinal cell in a retina comprising contacting the retina with thecompound of the compound of Formula (A), a compound that inhibits11-cis-retinol production with an IC50 of about 1 uM or less whenassayed in vitro, utilizing extract of cells that express RPE65 andLRAT, wherein the extract further comprises CRALBP, wherein the compoundis stable in solution for at least about 1 week at room temperature, ora non-retinoid compound that inhibits an isomerase reaction resulting inproduction of 11-cis retinol, wherein said isomerase reaction occurs inRPE, and wherein said compound has an ED50 value of 1 mg/kg or less whenadministered to a subject. In a further embodiment, the pharmaceuticalcomposition is administered under conditions and at a time sufficient toinhibit dark adaptation of a rod photoreceptor cell, thereby reducingischemia in the eye. In a specific embodiment is the method wherein theretinal cell is a retinal neuronal cell. In a certain embodiment, theretinal neuronal cell is a photoreceptor cell.

Thus, the compounds can be represented by Formula (I):

as an isolated E or Z stereoisomer or a mixture of E and Zstereoisomers, as a tautomer or a mixture of tautomers, or as apharmaceutically acceptable salt, hydrate, solvate, N-oxide or prodrugthereof, wherein:

R₁ and R₂ are each the same or different and independently hydrogen oralkyl;

R₃, R₄, R₅ and R₆ are each the same or different and independentlyhydrogen, halogen, —OR₁₂, alkyl or fluoroalkyl;

R₇ and R₈ are each the same or different and independently hydrogen oralkyl;

R₉ is hydrogen, alkyl, carbocyclyl or —C(═O)R₁₃;

R₁₀ is hydrogen or alkyl; or

R₉ and R₁₀, together with the nitrogen atom to which they are attached,form an N-heterocyclyl;

R₁₁ is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;

R₁₂ is hydrogen or alkyl;

R₁₃ is alkyl, carbocyclyl or aryl;

W is —C(R₁₄)(R₁₅)—, —O—, —S—, —S(═O)—, —S(═O)₂— or —N(R₁₂)—;

Y is —C(R₁₆)(R₁₇)—;

R₁₄ and R₁₅ are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR₁₂, —NR₁₈R₁₉ or carbocyclyl; or

R₁₄ and R₁₅ form an oxo;

R₁₆ and R₁₇ are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR₁₂, —NR₁₈R₁₉ or carbocyclyl; or

R₁₆ and R₁₇ form an oxo; or

R₁₄ and R₁₆ together form a direct bond to provide a double bondconnecting W and Y; or

R₁₄ and R₁₆ together form a direct bond, and R₁₅ and R₁₇ together form adirect bond to provide a triple bond connecting W and Y;

R₁₈ and R₁₉ are each the same or different and independently hydrogen,alkyl, carbocyclyl, or —C(═O)R₁₃,

provided that when R₁₁ is phenyl, the compound of Formula (I) is not:

-   2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]acetamide;-   (2S,3R)-amino-3-hydroxy-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)-ethenyl]phenyl]-butanamide;-   L-glutamic acid,    1-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]ester;-   glycine, 3-hydroxy-5-[(1E)-2(4-hydroxyphenyl)ethenyl]phenyl ester;-   (2S)-2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]propanamide;-   (2S)-2-amino-3-hydroxy-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]propanamide;-   (2S)-2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]-4-methyl-pentanamide;-   (2S)-2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]-3-methyl-butanamide;    or-   2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenylbutanamide.

In one embodiment, R₁ is hydrogen, and compound has a structure ofFormula (Ia):

as an isolated E or Z stereoisomer or a mixture of E and Zstereoisomers, as a tautomer or a mixture of tautomers, or as apharmaceutically acceptable salt, hydrate, solvate, N-oxide or prodrugthereof, wherein

R₂ is H or C₁-C₆ alkyl;

each of R₃, R₄, R₅, and R₆ is the same or different and independentlyhydrogen, C₁-C₆ alkyl, or C₁-C₆ alkoxy;

R₇ is H or C₁-C₆ alkyl;

R₉ is hydrogen, C₁-C₆ alkyl, —(CH₂)_(n)OH wherein n is 1-6, or—C(═O)R₁₃;

R₁₀ is hydrogen or C₁-C₆ alkyl; or

R₉ and R₁₀, together with the nitrogen atom to which they are attached,form an N-heterocyclyl;

W is —C(R₁₄)(R₁₅)—, —O—, —S—, —S(═O)—, —S(═O)₂— or —N(H)—;

Y is —C(R₁₆)(R₁₇)—;

R₁₂ is hydrogen or C₁-C₆ alkyl;

R₁₃ is C₁-C₆ alkyl;

R₁₄ and R₁₅ are each the same or different and independently hydrogen,halogen, C₁-C₆ alkyl, fluoroalkyl, —OR₁₂, —NH₂; or

R₁₄ and R₁₅ form an oxo;

R₁₆ and R₁₇ are each the same or different and independently hydrogen,halogen, C₁-C₆ alkyl, —OR₁₂;

or R₁₆ and R₁₇ form an oxo;

R₁₄ and R₁₆ together form a direct bond to provide a double bondconnecting W and Y; or

R₁₄ and R₁₆ together form a direct bond, and R₁₅ and R₁₇ together form adirect bond to provide a triple bond connecting W and Y; and

R₁₁ is alkyl; phenyl substituted with alkyl, —OR₁₂, —O(CH₂)_(n)OCH₃wherein n is 1-6, alkenyl, alkynyl, halogen, fluoroalkyl, phenyl, —SCH₃,or aralkyl; naphthenyl naphthenyl optionally substituted with alkyl,halogen, or —OR₁₂; carbocyclyl; cyclohexenyl optionally substituted withalkyl,

provided that R₁₁ is not 3,4,5-tri-methoxyphenyl.

In a specific embodiment, R₂ is hydrogen or n-butyl.

In certain specific embodiments, halogen is chloro or fluoro.

In other specific embodiments, each of R₃, R₄, R₅, and R₆ is the same ordifferent and independently hydrogen, methyl or methoxy. In certainspecific embodiments, at least two of R₃, R₄, R₅, and R₆ are hydrogen.In other certain specific embodiments, at least one of R₃, R₄, R₅, andR₆ is methyl or methoxy.

In a specific embodiment, R₉ is —(CH₂)_(n)OH wherein n is 2. In otherspecific embodiments, R₉ is —C(═O)R₁₃ wherein R₁₃ is methyl.

In another specific embodiment, each of R₉ and R₁₀ the same or differentand independently hydrogen or methyl. In other specific embodiments,each of R₉ and R₁₀ is hydrogen.

In other specific embodiments, when W is —C(R₁₄)(R₁₅)—, R₁₄ and R₁₅ areeach the same or different and independently hydrogen, fluoro, methyl,ethyl, trifluoromethyl, —OH, —OHCH₃, or —NH₂. In certain specificembodiments each of R₁₄ and R₁₅ is hydrogen.

In yet other specific embodiments Y is —CH₂—, —CH(CH₃)—, —CH(CH₃)₂—,—CHOH—, —CHF—, —CF₂—, or —C(═O)—.

In other specific embodiments, R₁₁ is phenyl substituted with alkyl,—OR₁₂, —O(CH₂)_(n)OCH₃ wherein n is 1-6, alkenyl, alkynyl, halogen,fluoroalkyl, phenyl, or —SCH₃. In other certain specific embodiments,halogen is chloro. In yet other specific embodiments, n is 2.

In other embodiments, R₁₁ is naphthenyl substituted with —OR₁₂, whereinR₁₂ is hydrogen or C₁-C₆ alkyl. In yet another specific embodiment, R₁₁is cyclohexenyl optionally substituted with C₁-C₆ alkyl; in anotherspecific embodiment, R₁₁ is tri-methyl-cyclohexenyl. In other specificembodiments, R₁₁ is alkyl optionally substituted with —OR₁₂ wherein R₁₂is hydrogen or C₁-C₆ alkyl.

One embodiment provides a compound having a structure of Formula (I)wherein R₁₁ is aryl or carbocyclyl, as defined herein.

A further embodiment provides a compound having a structure of Formula(I) wherein R₁₁ is aryl, which has a structure of Formula (II):

as an isolated E or Z stereoisomer or a mixture of E and Zstereoisomers, as a tautomer or a mixture of tautomers, or as apharmaceutically acceptable salt, hydrate, solvate, N-oxide or prodrugthereof, wherein:

R₁ and R₂ are each the same or different and independently hydrogen oralkyl;

R₃, R₄, R₅ and R₆ are each the same or different and independentlyhydrogen, halogen, —OR₁₂, alkyl or fluoroalkyl;

R₇ and R₈ are each the same or different and independently hydrogen oralkyl;

R₉ is hydrogen, alkyl, carbocyclyl or —C(═O)R₁₃;

R₁₀ is hydrogen or alkyl; or

R₉ and R₁₀, together with the nitrogen atom to which they are attached,form an N-heterocyclyl;

R₁₂ is hydrogen or alkyl;

R₁₃ is alkyl, carbocyclyl or aryl;

W is —C(R₁₄)(R₁₅)—, —S—, —S(═O)—, —S(═O)₂— or —N(R₁₂)—;

Y is —C(R₁₆)(R₁₇)—;

R₁₄ and R₁₅ are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR₁₂, —NR₁₈R₁₉ or carbocyclyl; or

R₁₄ and R₁₅ form an oxo;

R₁₆ and R₁₇ are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR₁₂, —NR₁₈R₁₉ or carbocyclyl; or

R₁₆ and R₁₇ form an oxo; or

R₁₄ and R₁₆ together form a direct bond to provide a double bondconnecting W and Y; or

R₁₄ and R₁₆ together form a direct bond, and R₁₅ and R₁₇ together form adirect bond to provide a triple bond connecting W and Y;

R₁₈ and R₁₉ are each the same or different and independently hydrogen,alkyl, carbocyclyl, or —C(═O)R₁₃,

t is 0, 1, 2, 3, 4 or 5; and

each R₂₀ is the same or different and independently alkyl, —OR₁₂,alkenyl, alkynyl, halo, fluoroalkyl, aryl or aralkyl, or

two adjacent R₂₀, together with the two carbon atoms to which they areattached, form a fused phenyl ring.

In one embodiment, each of R₉ and R₁₀ is hydrogen and the compound has astructure of Formula (IIa):

In one embodiment, W is —C(R₁₄)(R₁₅)— and the linkage —W—Y—C(R₇)(R₈)— isan alkylene chain. Thus, the compound has a structure of Formula (IIb):

as an isolated E or Z stereoisomer or a mixture of E and Zstereoisomers, as a tautomer or a mixture of tautomers, or as apharmaceutically acceptable salt, hydrate, solvate, N-oxide or prodrugthereof, wherein:

R₁ and R₂ are each the same or different and independently hydrogen oralkyl;

R₃, R₄, R₅ and R₆ are each the same or different and independentlyhydrogen, halogen, —OR₁₂, alkyl or fluoroalkyl;

R₇ and R₈ are each the same or different and independently hydrogen oralkyl;

R₁₂ is hydrogen or alkyl;

R₁₃ is alkyl, carbocyclyl or aryl;

R₁₄ and R₁₅ are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR₁₂, —NR₁₈R₁₉ or carbocyclyl; or

R₁₄ and R₁₅ form an oxo;

R₁₆ and R₁₇ are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR₁₂, —NR₁₈R₁₉ or carbocyclyl; or

R₁₆ and R₁₇ form an oxo; or

R₁₈ and R₁₉ are each the same or different and independently hydrogen,alkyl, carbocyclyl, or —C(═O)R₁₃,

t is 0, 1, 2, 3, 4 or 5; and

each R₂₀ is the same or different and independently alkyl, —OR₁₂,alkenyl, alkynyl, halo, fluoroalkyl, aryl or aralkyl, or

two adjacent R₂₀, together with the two carbon atoms to which they areattached, form a fused phenyl ring.

In certain embodiments, t is 0, 1, 2 or 3, each R₂₀ is independentlyalkyl, —OR₁₂, alkynyl, phenyl, halo or fluoroalkyl, and R₃, R₄, R₅ andR₆ are each independently hydrogen, alkyl, —OR₁₂, halo or fluoroalkyl.

In certain embodiments, R₇, R₈, R₁₄, R₁₅, R₁₆ and R₁₇ are eachindependently hydrogen, halogen, alkyl or —OR₁₂, wherein R₁₂ is hydrogenor alkyl.

In certain specific embodiments, the compounds of Formula (I), (II),(IIa) or (IIb) have the structures shown in Table 1. Each of thecompounds provided in Table 1 below and in the following Tables 2-11 asCompound Numbers may also be referred to herein as Example numbers. TheCompound number corresponds to the Example number herein that describesthe synthesis of the compound.

TABLE 1 Compound No. Compound Formula Chemical Name 1

(E)-3-(3-(2,6-dimethylstyryl)phenyl)propan-1-amine 2

(Z)-3-(3-(2,6-dimethylstyryl)phenyl)propan-1-amine 16

(E)-3-(3-(2-methylstyryl)phenyl)propan-1-amine 17

(Z)-3-(3-(2-methylstyryl)phenyl)propan-1-amine 28

(E)-3-(3-(2,6-dimethylstyryl)-2-methylphenyl)propan- 1-amine 29

(Z)-3-(3-(2,6-dimethylstyryl)-2-methylphenyl)propan- 1-amine 32

(E/Z)-3-(3-(2-ethyl-6-methylstyryl)phenyl)propan-1- amine 33

(E/Z)-3-(3-(2,5-dimethylstyryl)phenyl)propan-1-amine 34

(E/Z)-3-(3-(2,4-dimethylstyryl)phenyl)propan-1-amine 35

(E)-3-(3-(2,4,6-trimethylstyryl)phenyl)propan-1-amine 36

(E/Z)-3-(3-(2-ethylstyryl)phenyl)propan-1-amine 37

(E/Z)-3-(3-(2-ethynylstyryl)phenyl)propan-1-amine 38

(E/Z)-3-(3-(3,4-dimethylstyryl)phenyl)propan-1-amine 39

(E/Z)-3-(3-(2-isopropylstyryl)phenyl)propan-1-amine 40

(E/Z)-4-(3-(3,5-dimethylstyryl)phenyl)propan-1-amine 41

(E/Z)-4-(3-(2-methoxystyryl)phenyl)propan-1-amine 43

(E)-3-(3-(2,6-dichlorostyryl)phenyl)propan-1-amine 44

(E/Z)-3-(3-(2,3-dimethylstyryl)phenyl)propan-1-amine 46

(E)-3-(3-(2,6-dimethylstyryl)-4-fluorophenyl)propan- 1-amine 47

(E/Z)-3-(3-(2-(trifluoromethyl)styryl)phenyl)propan-1- amine 48

(E)-3-(3-(2,6-dimethoxystyryl)phenyl)propan-1-amine 49

(E)-3-(3-(2,6-bis(trifluoromethyl)styryl)- phenyl)propan-1-amine 50

(E)-3-amino-1-(3-(2,6-dichlorostyryl)phenyl)propan-1- 82

(E)-3-amino-1-(3-(2-chloro-6- methylstyryl)phenyl)propan-1-ol 70

(E)-2-(3-(3-aminopropyl)styryl)phenol 71

(E)-3-(5-(2,6-dichlorostyryl)-2- methoxyphenyl)propan-1-amine 74

(R,E)-1-amino-3-(3-(2,6-dichlorostyryl)phenyl)propan- 2-ol 75

(S,E)-1-amino-3-(3-(2,6-dichlorostyryl)phenyl)propan- 2-ol 76

(E/Z)-(3-(3-(2,6-diethoxystyryl)phenyl)propan-1- amine 77

(E)-3-(3-(2-ethoxystyryl)phenyl)propan-1-amine 78

(E/Z)-3-(3-(2-isopropoxystyryl)phenyl)propan-1-amine 80

(E)-3-amino-1-(3-(2,6-dichlorostyryl)phenyl)propan-1- one 81

(E)-1-amino-3-(3-(2,6-dichlorostyryl)phenyl)propan-2- one 86

(R,E)-3-amino-1-(3-(2,6-dichlorostyryl)phenyl)propan- 1-ol 87

(S,E)-3-amino-1-(3-(2,6-dichlorostyryl)phenyl)propan- 1-ol 88

(S,E)-3-(3-(2,6-dichlorostyryl)phenyl)-2-fluoropropan- 1-amine 89

(E)-3-(3-(2,6-dichlorostyryl)phenyl)-2,2- difluoropropan-1-amine 90

(Z)-3-(3-(2-(2-methoxyethoxy)styryl)phenyl)-propan- 1-amine 91

(E)-3-(3-(3-methoxystyryl)phenyl)propan-1-amine 93

(Z)-3-(3-(4-chlorostyryl)phenyl)propan-1-amine 94

(E)-3-(3-(2-(biphenyl-2-yl)vinyl)phenyl)propan-1- amine 97

(E)-3-(3-(3-chlorostyryl)phenyl)propan-1-amine 98

(E)-3-(3-(2-butoxystyryl)phenyl)propan-1-amine 99

(E)-3-(3-(4-methoxystyryl)phenyl)propan-1-amine 109

(Z)-3-(3-(2-Propoxystyryl)phenyl)propan-1-amine 107

(E)-3-(5-(2-Chloro-6-(methylthio)styryl)-2- methoxyphenyl)propan-1-amine110

(E)-3-(3-(2-phenylprop-1-enyl)phenyl)propan-1-amine

In certain embodiments, a compound of formula (IIb) is provided whereint is 2 or 3, two adjacent R₂₀, together with the two carbon atoms towhich they are attached, form a fused phenyl ring; optionally, a thirdR₂₀ is alkyl or —OR₁₂; and R₃, R₄, R₅ and R₆ are each independentlyhydrogen, alkyl, halo or fluoroalkyl.

In other certain embodiments, R₇, R₈, R₁₄, R₁₅, R₁₆ and R₁₇ are eachindependently hydrogen, halogen, alkyl or —OR₁₂.

In certain specific embodiments, the compounds of Formula (I), (II),(IIa) or (IIb) have a structure shown in Table 2.

TABLE 2 Compound No. Compound Formula Chemical Name 92

(E)-3-(3-(2-(1-methoxynaphthalen-2- yl)vinyl)phenyl)propan-1-amine 95

(Z)-3-(3-(2-(naphthalen-1-yl)vinyl)phenyl)propan-1- amine 96

(Z)-3-(3-(2-(3-methoxynaphthalen-2- yl)vinyl)phenyl)propan-1-amine 108

(E/Z)-3-(3-(2-(2-methoxynaphthalen-1- yl)vinyl)phenyl)propan-1-amine

Another embodiment provides a compound having a structure of Formula(IIa) wherein W is —O—, —S—, —S(═O)—, —S(═O)₂— or —N(R₁₂)—.

In certain embodiments, t is 0, 1, 2 or 3, each R₂₀ is independentlyalkyl, —OR₁₂ or halo, and R₃, R₄, R₅ and R₆ are each independentlyhydrogen, alkyl or halo. In certain specific embodiments, thesecompounds have a structure shown in Table 3.

TABLE 3 Compound No. Chemical Formula Chemical Name 15

(E)-2-amino-N-(3-(2,6-dimethylstyryl)phenyl)acet- amide 18

(E)-2-(3-(2,6-dimethylstyryl)phenylthio)ethanamine 19

(E)-2-(3-(2,6-dimethylstyryl)phenyl- sulfinyl)ethanamine 20

(E)-2-(3-(2,6-dimethylstyryl)phenyl- sulfonyl)ethanamine 119

(S,E)-1-(3-(1-aminopropan-2- yloxy)styryl)cyclohexanol 112

(E)-1-(3-(2-aminoethoxy)styryl)cyclohexanol

Another embodiment provides a compound having a structure of Formula(IIa) wherein W and Y are connected by a double bond. One exemplarycompound of this embodiment is shown in Table 3A:

TABLE 3A Compound No. Chemical Formula Chemical Name 45

(E/Z)-3-(3-(2,6-Dimethylstyryl)phenyl)prop-2-en-1- amine

Another embodiment provides a compound having a structure of Formula(II), wherein R₉ and R₁₀ together with the nitrogen to which they areattached form a N-heterocyclyl. In certain embodiments, theN-heterocyclyl is morpholinyl, pyrrolidinyl, piperidinyl or piperazinyl.

In other certain embodiments, each of R₁ and R₂ is hydrogen, t is 0, 1,2 or 3, each R₂₀ is independently alkyl or halo, and R₃, R₄, R₅ and R₆are each independently hydrogen, alkyl or halo.

In certain embodiments, W is —C(R₁₄)(R₁₅)— and the linkage—W—Y—C(R₇)(R₈)— is an alkylene chain. In certain specific embodiments,exemplary compounds have a structure shown in Table 4.

TABLE 4 Compound No. Chemical Formula Chemical Name 3

(E)-4-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)morpholine 4

(Z)-4-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)morpholine 5

(E)-1-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)pyrrolidine 6

(Z)-1-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)pyrrolidine 11

(E)-1-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)piperidine 12

(Z)-1-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)piperidine 13

(E)-1-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)piperazine 14

(Z)-1-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)piperazine

Another embodiment provides a compound having a structure of Formula(II), wherein R₉ is alkyl or —C(═O)R₁₃, wherein R₁₃ is alkyl, and R₁₀ ishydrogen or alkyl.

In certain embodiments, each of R₁ and R₂ is hydrogen, t is 0, 1, 2 or3, each R₂₀ is independently alkyl or halo, and R₃, R₄, R₅ and R₆ areeach independently hydrogen, alkyl or halo.

In yet other certain embodiments, W is —C(R₁₄)(R₁₅)— and the linkage—W—Y—C(R₇)(R₈)— is an alkylene chain. In specific embodiments, thecompound has a structure shown in Table 5.

TABLE 5 Compound No. Chemical Formula Chemical Name 7

(E)-3-(3-(2,6-dimethylstyryl)phenyl)-N-methylpropan- 1-amine 8

(Z)-3-(3-(2,6-dimethylstyryl)phenyl)-N-methylpropan-1- amine 9

(E)-3-(3-(2,6-dimethylstyryl)phenyl)-N,N- dimethylpropan-1-amine 10

(Z)-3-(3-(2,6-dimethylstyryl)phenyl)-N,N- dimethylpropan-1-amine 51

(E/Z)-N-(3-(3-(2,6-dimethylstyryl)phenyl)prop- yl)acetamide 52

(E/Z)-N-(3-(3-(2,6-dimethylstyryl)phenyl)prop- yl)pentadecanamide

A further embodiment provides a compound having a structure of Formula(I) wherein R₁₁ is carbocyclyl, as defined herein.

In certain embodiments, R₁₁ is 5-, 6-, or 7-member cycloalkyl ring.

In certain specific embodiments, R₁₁ is cyclohexyl. One exemplarycompound of this embodiment is shown in Table 5A:

TABLE 5A Compound No. Chemical Formula Chemical Name 22

(E)-3-(3-(2-cyclohexylvinyl)phenyl)propan-1-amine

In another embodiment, R₁₁ is a 5-, 6-, or 7-member cycloalkenyl.

In yet other embodiments, R₁₁ is cyclohexenyl and the compound has astructure of Formula (III):

as an isolated E or Z stereoisomer or a mixture of E and Zstereoisomers, as a tautomer or a mixture of tautomers, or as apharmaceutically acceptable salt, hydrate, solvate, N-oxide or prodrugthereof, wherein:

R₁ and R₂ are each the same or different and independently hydrogen oralkyl;

R₃, R₄, R₅ and R₆ are each the same or different and independentlyhydrogen, halogen, —OR₁₂, alkyl or fluoroalkyl;

R₇ and R₈ are each the same or different and independently hydrogen oralkyl;

R₉ is hydrogen, alkyl, carbocyclyl or —C(═O)R₁₃;

R₁₀ is hydrogen or alkyl; or

R₉ and R₁₀, together with the nitrogen atom to which they are attached,form an N-heterocyclyl;

R₁₂ is hydrogen or alkyl;

R₁₃ is alkyl, carbocyclyl or aryl;

W is —C(R₁₄)(R₁₅)—, —O—, —S—, —S(═O)—, —S(═O)₂— or —N(R₁₂)—;

Y is —C(R₁₆)(R₁₇)—;

R₁₄ and R₁₅ are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR₁₂, —NR₁₈R₁₉ or carbocyclyl; or

R₁₄ and R₁₅ form an oxo;

R₁₆ and R₁₇ are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR₁₂, —NR₁₈R₁₉ or carbocyclyl; or

R₁₆ and R₁₇ form an oxo; or

R₁₄ and R₁₆ together form a direct bond to provide a double bondconnecting W and Y; or

R₁₄ and R₁₆ together form a direct bond, and R₁₅ and R₁₇ together form adirect bond to provide a triple bond connecting W and Y;

R₁₈ and R₁₉ are each the same or different and independently hydrogen,alkyl, carbocyclyl, or —C(═O)R₁₃,

p is 0, 1, 2, 3, 4, 5, 7, 8 or 9; and

each R₂₁ is the same or different and independently alkyl, —OR₁₂,alkenyl, alkynyl, halo, fluoroalkyl or aralkyl.

In certain embodiments, W is —C(R₁₄)(R₁₅)— and the linkage—W—Y—C(R₇)(R₈)— is an alkylene chain. Thus, the compound has a structureof Formula (IIIa):

as an isolated E or Z stereoisomer or a mixture of E and Zstereoisomers, as a tautomer or a mixture of tautomers, or as apharmaceutically acceptable salt, hydrate, solvate, N-oxide or prodrugthereof,

wherein:

R₁ and R₂ are each the same or different and independently hydrogen oralkyl;

R₃, R₄, R₅ and R₆ are each the same or different and independentlyhydrogen, halogen, —OR₁₂, alkyl or fluoroalkyl;

R₇ and R₈ are each the same or different and independently hydrogen oralkyl;

R₉ is hydrogen, alkyl, carbocyclyl or —C(═O)R₁₃;

R₁₀ is hydrogen or alkyl; or

R₉ and R₁₀, together with the nitrogen atom to which they are attached,form an N-heterocyclyl;

R₁₂ is hydrogen or alkyl;

R₁₃ is alkyl, carbocyclyl or aryl;

R₁₄ and R₁₅ are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR₁₂, —NR₁₈R₁₉ or carbocyclyl; or

R₁₄ and R₁₅ form an oxo;

R₁₆ and R₁₇ are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR₁₂, —NR₁₈R₁₉ or carbocyclyl; or

R₁₆ and R₁₇ form an oxo; or

R₁₄ and R₁₆ together form a direct bond to provide a double bondconnecting W and Y; or

R₁₄ and R₁₆ together form a direct bond, and R₁₅ and R₁₇ together form adirect bond to provide a triple bond connecting W and Y;

R₁₈ and R₁₉ are each the same or different and independently hydrogen,alkyl, carbocyclyl, or —C(═O)R₁₃,

p is 0, 1, 2, 3, 4, 5, 7, 8 or 9; and

each R₂₁ is the same or different and independently alkyl, —OR₁₂,alkenyl, alkynyl, halo, fluoroalkyl or aralkyl.

One embodiment provides a compound having a structure of Formula (IIIa)wherein each of R₉ and R₁₀ is hydrogen.

In certain embodiments, each of R₁ and R₂ is hydrogen, p is 0, 1, 2 or3, each R₂₁ is independently alkyl, halo or fluoroalkyl, and each of R₃,R₄, R₅ and R₆ is independently hydrogen, alkyl, halo, fluoroalkyl or—OR₁₂.

In other certain embodiments, R₇, R₈, R₁₄, R₁₅, R₁₆ and R₁₇ are eachindependently hydrogen, halogen, alkyl, fluoroalkyl, —OR₁₂ or —NR₁₈R₁₉,wherein R₁₂ is hydrogen or alkyl, and R₁₈ and R₁₉ are each independentlyhydrogen or alkyl. In certain specific embodiments, a compound offormula (I), (III) or (IIIa) has a structure as shown in Table 6.

TABLE 6 Compound No. Chemical Formula Chemical Name 21

(E)-3-(3-(2-(2,6,6-trimethylcyclohex-1- enyl)vinyl)phenyl)propan-1-amine25

(E)-3-amino-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-ol 105

(S,E)-3-amino-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-ol 106

(R,E)-3-Amino-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-ol 26

(E)-2-methyl-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine 27

(E)-3-(3-(2-(2,6,6-trimethylcyclohex-1- enyl)vinyl)phenyl)butan-1-amine53

(E)-3-(2-methyl-5-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine 54

(E/Z)-4-amino-2-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-2-ol 55

(E)-3-fluoro-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine 57

(E)-4-amino-1,1,1-trifluoro-2-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-2-ol 58

(E)-3-amino-2,2-dimethyl-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-ol 59

(E)-3-amino-2-methyl-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-ol 61

(E)-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propane-1,3-diamine 62

(E)-4,4,4-trifluoro-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-1-amine 64

(E)-3-methoxy-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine 65

(E)-4-(3-(2-(2,6,6-trimethylcyclohex-1- enyl)vinyl)phenyl)butan-2-amine66

(E)-1-amino-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-2-ol 111

(E)-1-(3-(3-amino-1-hydroxypropyl)styryl)cyclohexanol 113

(E)-1-(3-((1R,2R)-3-amino-1-hydroxy-2- methylpropyl)styryl)cyclohexanol114

(E)-1-(3-(3-amino-1-hydroxypropyl)-5- fluorostyryl)cyclohexanol 115

(E)-1-(3-(3-amino-1-hydroxypropyl)-2- fluorostyryl)cyclohexanol 116

(E)-4-(3-(3-amino-1-hydroxypropyl)styryl)heptan-4-ol 117

(1S,2S)-3-amino-1-(3-((E)-2-(1-hydroxycyclohexyl)vinyl)phenyl)propane-1,2-diol 118

(1R,2R)-3-amino-1-(3-((E)-2-(1-hydroxycyclohexyl)vinyl)phenyl)propane-1,2-diol 120

(E)-1-(5-(3-amino-1-hydroxypropyl)-2- methoxystyryl)cyclohexanol 121

(E)-1-(3-(3-amino-1-hydroxypropyl)-4- chlorostyryl)cyclohexanol 122

(E)-1-(3-(3-amino-1-hydroxypropyl)-4- methylstyryl)cyclohexanol 123

(E)-1-(3-(3-amino-1-hydroxypropyl)-5- methylstyryl)cyclohexanol 124

(1S,2R)-3-amino-1-(3-((E)-2-(1-hydroxycyclohexyl)vinyl)phenyl)propane-1,2-diol 125

(E)-2-(3-(3-amino-1-hydroxypropyl)styryl)cyclohexanol 126

(E)-1-(5-(3-amino-1-hydroxypropyl)-2- fluorostyryl)cyclohexanol 127

(E)-1-(3-(3-amino-1-hydroxypropyl)-5- methoxystyryl)cyclohexanol 128

(E)-1-(3-(3-amino-1-hydroxypropyl)-4- fluorostyryl)cyclohexanol 129

(1R,2S)-3-amino-1-(3-((E)-2-(1-hydroxycyclohexyl)vinyl)phenyl)propane-1,2-diol 130

(E)-1-(3-(3-amino-1-hydroxypropyl)-5- chlorostyryl)cyclohexanol 131

(E)-1-(3-(3-amino-1-hydroxypropyl)-2- methoxystyryl)cyclohexanol

Another embodiment provides a compound having a structure of Formula(IIIa) wherein R₉ is alkyl and R₁₀ is hydrogen.

In certain embodiments, each of R₁ and R₂ is hydrogen, p is 0, 1, 2 or3, each R₂₁ is independently alkyl, halo or fluoroalkyl, and each of R₃,R₄, R₅ and R₆ is independently hydrogen, alkyl, halo, fluoroalkyl or—OR₁₂.

In certain embodiments, R₇, R₈, R₁₄, R₁₅, R₁₆ and R₁₇ are eachindependently hydrogen, halogen, alkyl, fluoroalkyl or —OR₁₂, whereinR₁₂ is hydrogen or alkyl. In certain specific embodiments, the compoundof formula (III) and (IIIa) has a structure shown in Table 7.

TABLE 7 Compound No. Chemical Formula Chemical Name 42

(E)-2-(3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propylamino)ethanol 63

(E)-3-methoxy-N-methyl-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine

Another embodiment provides a compound of Formula (IIIa), wherein R₇,R₈, R₁₆ and R₁₇ are each independently hydrogen, halogen, alkyl,fluoroalkyl or —OR₁₂, wherein R₁₂ is hydrogen or alkyl, and R₁₄ and R₁₅together form oxo. In certain specific embodiments, a compound offormula (III) and (IIIa) has a structure shown in Table 8.

TABLE 8 Compound No. Chemical Formula Chemical Name 56

(E)-3-amino-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-one 60

(E)-3-amino-2,2-dimethyl-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-one 72

(E)-3-amino-2-methyl-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-one 73

(E)-3-amino-2-fluoro-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-one

A further embodiment provides a compound having a structure of Formula(III) wherein W is —NH— or —O—.

In one embodiment, each of R₁, R₂, R₉ and R₁₀ is hydrogen.

In certain embodiments, p is 0, 1, 2 or 3, each R₂₁ is independentlyalkyl or halo, and R₃, R₄, R₅ and R₆ are each independently hydrogen,alkyl, halo or fluoroalkyl. In certain embodiments, the compounds ofFormula (I) or (III) are those shown in Table 9.

TABLE 9 Compound No. Chemical Formula Chemical Name 31

(E)-2-(3-(2-(2,6,6-trimethylcyclohex-1- enyl)vinyl)phenoxy)ethanamine 30

(E)-2-amino-N-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)acetamide

A further embodiment provides a compound having a structure of Formula(III) wherein W and Y are connected by a double or triple bond.

In one embodiment, each of R₁, R₂, R₉ and R₁₀ is hydrogen.

In certain embodiments, p is 0, 1, 2 or 3, each R₂₁ is independentlyalkyl or halo, R₃, R₄, R₅ and R₆ are each independently hydrogen, alkyl,halo or fluoroalkyl, and R₁₅ and R₁₇ are each independently hydrogen,alkyl or halogen. In certain specific embodiments, a compound of Formula(I) or (III) has a structure shown in Table 10.

TABLE 10 Compound No. Chemical Formula Chemical Name 68

(E)-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)prop-2-yn-1-amine 67

(E)-2-fluoro-3-(3-((E)-2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)prop-2-en-1-amine

A further embodiment provides a compound having a structure of Formula(I) wherein R₁₁ is alkyl.

In one embodiment, each of R₉ and R₁₀ is hydrogen.

In certain embodiments, W is —C(R₁₄)(R₁₅)— and the linkage—W—Y—C(R₇)(R₈)— is an alkylene chain. In certain embodiments, R₁, R₂,R₃, R₄, R₅ and R₆ are each independently hydrogen or alkyl.

In certain specific embodiments, a compound of Formula (I) wherein R₁₁is alkyl has a structure shown in Table 11.

TABLE 11 Compound No. Chemical Formula Chemical Name 23

(E)-3-(3-(pent-1-enyl)phenyl)propan-1-amine 24

(E)-3-(3-(hept-1-enyl)phenyl)propan-1-amine 69

(E)-3-(3-(non-4-en-5-yl)phenyl)propan-1-amine 79

(E)-4-(3-(3-aminopropyl)phenyl)-2-methylbut-3-en-2-ol 83

(E)-4-(3-(3-aminopropyl)styryl)heptan-4-ol 84

(E)-1-(3-(3-aminopropyl)phenyl)hex-1-en-3-ol 85

(E)-4-(3-(2-aminoethoxy)styryl)heptan-4-ol 100

(E)-1-(3-(3-aminopropyl)phenyl)-3-ethylpent-1-en-3-ol 101

(E)-3-(3-(3-aminopropyl)phenyl)prop-2-en-1-ol 102

(E)-3-(3-(3-methoxyprop-1-enyl)phenyl)propan-1-amine - 103

(E)-1-(3-(3-aminopropyl)phenyl)-3-methylhex-1-en-3-ol 104

(E)-1-(3-(3-aminopropyl)phenyl)-3-ethylhex-1-en-3-ol

In certain specific embodiments, a compound of Formula (A) has astructure shown in Table 12.

TABLE 12 132

(E)-4-(3-(2,6-Dimethylstyryl)phenyl)butan-1-amine 133

1-(3-(2,6-Dimethylstyryl)phenyl)-N,N- dimethylmethanamine 134

4-(3-(2,6-Dimethylstyryl)benzyl)morpholine 135

(E)-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl) methanamine 136

(E)-(3-(2-(2,6,6-trimethylcyclohex-1- enyl)vinyl)phenyl)methanamine 137

(E)-(3-(2-(2,6,6-trimethylcyclohex-1- enyl)vinyl)phenyl)methanamine

Definitions

As used in the specification and appended claims, unless specified tothe contrary, the following terms have the meaning indicated:

As used herein and in the appended claims, the singular forms “a,”“and,” and “the” include plural referents unless the context clearlydictates otherwise. Thus, for example, reference to “a compound”includes a plurality of such compounds, and reference to “the cell”includes reference to one or more cells (or to a plurality of cells) andequivalents thereof known to those skilled in the art, and so forth.When ranges are used herein for physical properties, such as molecularweight, or chemical properties, such as chemical formulae, allcombinations and subcombinations of ranges and specific embodimentstherein are intended to be included. The term “about” when referring toa number or a numerical range means that the number or numerical rangereferred to is an approximation within experimental variability (orwithin statistical experimental error), and thus the number or numericalrange may vary between 1% and 15% of the stated number or numericalrange. The term “comprising” (and related terms such as “comprise” or“comprises” or “having” or “including”) is not intended to exclude thatin other certain embodiments, for example, an embodiment of anycomposition of matter, composition, method, or process, or the like,described herein, may “consist of” or “consist essentially of” thedescribed features.

“Amino” refers to the —NH₂ radical.

“Cyano” refers to the —CN radical.

“Nitro” refers to the —NO₂ radical.

“Oxa” refers to the —O— radical.

“Oxo” refers to the ═O radical.

“Oximo” refers to the ═N—OH radical.

“Imino” refers to the ═N—H radical.

“Hydrazino” refers to the ═N—NH₂ radical.

“Thioxo” refers to the ═S radical.

“Alkyl” refers to a straight or branched hydrocarbon chain radicalconsisting solely of carbon and hydrogen atoms, containing nounsaturation, having from one to fifteen carbon atoms. In certainembodiments, an alkyl comprises one to eight carbon atoms. In otherembodiments, an alkyl comprises one to six carbon atoms. The alkyl isattached to the rest of the molecule by a single bond, for example,methyl (Me), ethyl (Et), n-propyl, 1-methylethyl (iso-propyl), n-butyl,n-pentyl, 1,1-dimethylethyl (t-butyl), 3-methylhexyl, 2-methylhexyl, andthe like. Unless stated otherwise specifically in the specification, analkyl group is optionally substituted by one or more of the followingsubstituents: halo, cyano, nitro, oxo, thioxo, trimethylsilanyl,—OR^(a), —SR^(a), —OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a),—C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a),—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2) and —S(O)_(t)N(R^(a))₂ (where t is 1 or 2) where each R^(a) isindependently hydrogen, alkyl, fluoroalkyl, carbocyclylcarbocyclylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl,heteroaryl or heteroarylalkyl.

“Alkenyl” refers to a straight or branched hydrocarbon chain radicalgroup consisting solely of carbon and hydrogen atoms, containing atleast one double bond, and having from two to twelve carbon atoms. Incertain embodiments, an alkenyl comprises two to eight carbon atoms. Inother embodiments, an alkenyl comprises two to four carbon atoms. Thealkenyl is attached to the rest of the molecule by a single bond, forexample, ethenyl (i.e., vinyl), prop-1-enyl (i.e., allyl), but-1-enyl,pent-1-enyl, penta-1,4-dienyl, and the like. Unless stated otherwisespecifically in the specification, an alkenyl group is optionallysubstituted by one or more of the following substituents: halo, cyano,nitro, oxo, thioxo, trimethylsilanyl, —OR^(a), —SR^(a), —OC(O)—R^(a),—N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a), —C(O)N(R^(a))₂,—N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a), —N(R^(a))S(O)_(t)R^(a) (where tis 1 or 2), —S(O)_(t)OR^(a) (where t is 1 or 2) and —S(O)_(t)N(R^(a))₂(where t is 1 or 2) where each R^(a) is independently hydrogen, alkyl,fluoroalkyl, carbocyclyl carbocyclylalkyl, aryl, aralkyl, heterocyclyl,heterocyclylalkyl, heteroaryl or heteroarylalkyl.

“Alkynyl” refers to a straight or branched hydrocarbon chain radicalgroup consisting solely of carbon and hydrogen atoms, containing atleast one triple bond, having from two to twelve carbon atoms. Incertain embodiments, an alkynyl comprises two to eight carbon atoms. Inother embodiments, an alkynyl has two to four carbon atoms. The alkynylis attached to the rest of the molecule by a single bond, for example,ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like. Unlessstated otherwise specifically in the specification, an alkynyl group isoptionally substituted by one or more of the following substituents:halo, cyano, nitro, oxo, thioxo, trimethylsilanyl, —OR^(a), —SR^(a),—OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a), —C(O)N(R^(a))₂,—N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a), —N(R^(a))S(O)_(t)R^(a) (where tis 1 or 2), —S(O)_(t)OR^(a) (where t is 1 or 2) and —S(O)_(t)N(R^(a))₂(where t is 1 or 2) where each R^(a) is independently hydrogen, alkyl,fluoroalkyl, carbocyclyl carbocyclylalkyl, aryl, aralkyl, heterocyclyl,heterocyclylalkyl, heteroaryl or heteroarylalkyl.

“Alkylene” or “alkylene chain” refers to a straight or branched divalenthydrocarbon chain linking the rest of the molecule to a radical group,consisting solely of carbon and hydrogen, containing no unsaturation andhaving from one to twelve carbon atoms, for example, methylene,ethylene, propylene, n-butylene, and the like. The alkylene chain isattached to the rest of the molecule through a single bond and to theradical group through a single bond. The points of attachment of thealkylene chain to the rest of the molecule and to the radical group canbe through one carbon in the alkylene chain or through any two carbonswithin the chain. Unless stated otherwise specifically in thespecification, an alkylene chain is optionally substituted by one ormore of the following substituents: halo, cyano, nitro, aryl,cycloalkyl, heterocyclyl, heteroaryl, oxo, thioxo, trimethylsilanyl,—OR^(a), —SR^(a), —OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a),—C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a),—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2) and —S(O)_(t)N(R^(a))₂ (where t is 1 or 2) where each R^(a) isindependently hydrogen, alkyl, fluoroalkyl, carbocyclylcarbocyclylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl,heteroaryl or heteroarylalkyl.

“Alkenylene” or “alkenylene chain” refers to a straight or brancheddivalent hydrocarbon chain linking the rest of the molecule to a radicalgroup, consisting solely of carbon and hydrogen, containing at least onedouble bond and having from two to twelve carbon atoms, for example,ethenylene, propenylene, n-butenylene, and the like. The alkenylenechain is attached to the rest of the molecule through a double bond or asingle bond and to the radical group through a double bond or a singlebond. The points of attachment of the alkenylene chain to the rest ofthe molecule and to the radical group can be through one carbon or anytwo carbons within the chain. Unless stated otherwise specifically inthe specification, an alkenylene chain is optionally substituted by oneor more of the following substituents: halo, cyano, nitro, aryl,cycloalkyl, heterocyclyl, heteroaryl, oxo, thioxo, trimethylsilanyl,—OR^(a), SR^(a), —OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a),—C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a),—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2) and —S(O)_(t)N(R^(a))₂ (where t is 1 or 2) where each R^(a) isindependently hydrogen, alkyl, fluoroalkyl, cycloalkyl, cycloalkylalkyl,aryl (optionally substituted with one or more halo groups), aralkyl,heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl, andwhere each of the above substituents is unsubstituted unless otherwiseindicated.

“Aryl” refers to a radical derived from an aromatic monocyclic ormulticyclic hydrocarbon ring system by removing a hydrogen atom from aring carbon atom. The aromatic monocyclic or multicyclic hydrocarbonring system contains only hydrogen and carbon from six to eighteencarbon atoms, where at least one of the rings in the ring system isfully unsaturated, i.e., it contains a cyclic, delocalized (4n+2)π-electron system in accordance with the Hückel theory. Aryl groupsinclude, but are not limited to, groups such as phenyl, fluorenyl, andnaphthyl. Unless stated otherwise specifically in the specification, theterm “aryl” or the prefix “ar-” (such as in “aralkyl”) includes arylradicals optionally substituted by one or more substituentsindependently selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl,cyano, nitro, optionally substituted aryl, optionally substitutedaralkyl, optionally substituted aralkenyl, optionally substitutedaralkynyl, optionally substituted carbocyclyl, optionally substitutedcarbocyclylalkyl, optionally substituted heterocyclyl, optionallysubstituted heterocyclylalkyl, optionally substituted heteroaryl,optionally substituted heteroarylalkyl, R^(b) OR^(a), R^(b) SR^(a),—R^(b)—OC(O)—R^(a), —R^(b)—N(R^(a))₂, —R^(b)—C(O)R^(a),—R^(b)—C(O)OR^(a), —R^(b)—C(O)N(R^(a))₂, —R^(b)—O—R^(c)—C(O)N(R^(a))₂,—R^(b)—N(R^(a))C(O)OR^(a), —R^(b)—N(R^(a))C(O)R^(a),—R^(b)—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —R^(b)—S(O)_(t)OR^(a)(where t is 1 or 2) and —R^(b)—S(O)_(t)N(R^(a))₂ (where t is 1 or 2),where each R^(a) is independently hydrogen, alkyl, fluoroalkyl,cycloalkyl, cycloalkylalkyl, aryl (optionally substituted with one ormore halo groups), aralkyl, heterocyclyl, heterocyclylalkyl, heteroarylor heteroarylalkyl, each R^(b) is independently a direct bond or astraight or branched alkylene or alkenylene chain, and R^(c) is astraight or branched alkylene or alkenylene chain, and where each of theabove substituents is unsubstituted unless otherwise indicated.

“Aralkyl” refers to a radical of the formula —R^(c)-aryl where R^(c) isan alkylene chain as defined above, for example, benzyl, diphenylmethyland the like. The alkylene chain part of the aralkyl radical isoptionally substituted as described above for an alkylene chain. Thearyl part of the aralkyl radical is optionally substituted as describedabove for an aryl group.

“Aralkenyl” refers to a radical of the formula —R^(d)-aryl where R^(d)is an alkenylene chain as defined above. The aryl part of the aralkenylradical is optionally substituted as described above for an aryl group.The alkenylene chain part of the aralkenyl radical is optionallysubstituted as defined above for an alkenylene group.

“Aralkynyl” refers to a radical of the formula —R^(e)-aryl, where R^(e)is an alkynylene chain as defined above. The aryl part of the aralkynylradical is optionally substituted as described above for an aryl group.The alkynylene chain part of the aralkynyl radical is optionallysubstituted as defined above for an alkynylene chain.

“Carbocyclyl” refers to a stable non-aromatic monocyclic or polycyclichydrocarbon radical consisting solely of carbon and hydrogen atoms,which include fused or bridged ring systems, having from three tofifteen carbon atoms. In certain embodiments, a carbocyclyl comprisesthree to ten carbon atoms. In other embodiments, a carbocyclyl comprisesfive to seven carbon atoms. The carbocyclyl is attached to the rest ofthe molecule by a single bond. Carbocyclyl is saturated, (i.e.,containing single C—C bonds only) or unsaturated (i.e., containing oneor more double bonds or triple bonds.) A fully saturated carbocyclylradical is also referred to as “cycloalkyl.” Examples of monocycliccycloalkyls include, e.g., cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl, and cyclooctyl. An unsaturated carbocyclyl isalso referred to as “cycloalkenyl.” Examples of monocyclic cycloalkenylsinclude, e.g., cyclopentenyl, cyclohexenyl, cycloheptenyl, andcyclooctenyl. Polycyclic carbocyclyl radicals include, for example,adamantyl, norbornyl (i.e., bicyclo[2.2.1]heptanyl), norbornenyl,decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. Unlessotherwise stated specifically in the specification, the term“carbocyclyl” includes carbocyclyl radicals that are optionallysubstituted by one or more substituents independently selected fromalkyl, alkenyl, alkynyl, halo, fluoroalkyl, oxo, thioxo, cyano, nitro,optionally substituted aryl, optionally substituted aralkyl, optionallysubstituted aralkenyl, optionally substituted aralkynyl, optionallysubstituted carbocyclyl, optionally substituted carbocyclylalkyl,optionally substituted heterocyclyl, optionally substitutedheterocyclylalkyl, optionally substituted heteroaryl, optionallysubstituted heteroarylalkyl, R^(b) OR^(a), —R^(b)—SR^(a),—R^(b)—OC(O)—R^(a), —R^(b)—N(R^(a))₂, —R^(b)—C(O)R^(a),—R^(b)—C(O)OR^(a), —R^(b)—C(O)N(R^(a))₂, —R^(b)—O—R^(c)—C(O)N(R^(a))₂,—R^(b)—N(R^(a))C(O)OR^(a), —R^(b)—N(R^(a))C(O)R^(a),—R^(b)—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —R^(b)—S(O)_(t)OR^(a)(where t is 1 or 2) and —R^(b)—S(O)_(t)N(R^(a))₂ (where t is 1 or 2),where each R^(a) is independently hydrogen, alkyl, fluoroalkyl,cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl,heterocyclylalkyl, heteroaryl or heteroarylalkyl, each R^(b) isindependently a direct bond or a straight or branched alkylene oralkenylene chain, and R^(c) is a straight or branched alkylene oralkenylene chain, and where each of the above substituents isunsubstituted unless otherwise indicated.

“Carbocyclylalkyl” refers to a radical of the formula —R^(c)-carbocyclylwhere R^(c) is an alkylene chain as defined above. The alkylene chainand the carbocyclyl radical is optionally substituted as defined above.

“Halo” or “halogen” refers to bromo, chloro, fluoro or iodo.

“Fluoroalkyl” refers to an alkyl radical, as defined above, that issubstituted by one or more fluoro radicals, as defined above, forexample, trifluoromethyl, difluoromethyl, 2,2,2-trifluoroethyl,1-fluoromethyl-2-fluoroethyl, and the like. The alkyl part of thefluoroalkyl radical is optionally substituted as defined above for analkyl group.

“Heterocyclyl” refers to a stable 3- to 18-membered non-aromatic ringradical that comprises two to twelve carbon atoms and from one to sixheteroatoms selected from nitrogen, oxygen and sulfur. Unless statedotherwise specifically in the specification, the heterocyclyl radical isa monocyclic, bicyclic, tricyclic or tetracyclic ring system, whichincludes fused or bridged ring systems. The heteroatoms in theheterocyclyl radical are optionally oxidized. One or more nitrogenatoms, if present, are optionally quaternized. The heterocyclyl radicalis partially or fully saturated. The heterocyclyl is attached to therest of the molecule through any atom of the ring(s). Examples of suchheterocyclyl radicals include, but are not limited to, dioxolanyl,thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl,imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl,octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl,2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl,piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl,thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl,thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and1,1-dioxo-thiomorpholinyl. Unless stated otherwise specifically in thespecification, the term “heterocyclyl” includes heterocyclyl radicals asdefined above that are optionally substituted by one or moresubstituents selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl,oxo, thioxo, cyano, nitro, optionally substituted aryl, optionallysubstituted aralkyl, optionally substituted aralkenyl, optionallysubstituted aralkynyl, optionally substituted carbocyclyl, optionallysubstituted carbocyclylalkyl, optionally substituted heterocyclyl,optionally substituted heterocyclylalkyl, optionally substitutedheteroaryl, optionally substituted heteroarylalkyl, —R^(b)—OR^(a),—R^(b)—SR^(a), —R^(b)—OC(O)—R^(a), —R^(b)—N(R^(a))₂, —R^(b)—C(O)R^(a),—R^(b)—C(O)OR^(a), —R^(b)—C(O)N(R^(a))₂, —R^(b)—O—R^(c)—C(O)N(R^(a))₂,—R^(b)—N(R^(a))C(O)OR^(a), —R^(b)—N(R^(a))C(O)R^(a),—R^(b)—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —R^(b)—S(O)_(t)OR^(a)(where t is 1 or 2) and —R^(b)—S(O)_(t)N(R^(a))₂ (where t is 1 or 2),where each R^(a) is independently hydrogen, alkyl, fluoroalkyl,cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl,heterocyclylalkyl, heteroaryl or heteroarylalkyl, each R^(b) isindependently a direct bond or a straight or branched alkylene oralkenylene chain, and R^(c) is a straight or branched alkylene oralkenylene chain, and where each of the above substituents isunsubstituted unless otherwise indicated.

“N-heterocyclyl” refers to a heterocyclyl radical as defined abovecontaining at least one nitrogen and where the point of attachment ofthe heterocyclyl radical to the rest of the molecule is through anitrogen atom in the heterocyclyl radical. An N-heterocyclyl radical isoptionally substituted as described above for heterocyclyl radicals.Examples of such N-heterocyclyl radicals include, but are not limitedto, morpholinyl, piperidinyl, piperazinyl, pyrrolidinyl, pyrazolidinyl,imidazolinyl, and imidazolidinyl.

“Heterocyclylalkyl” refers to a radical of the formula—R^(c)-heterocyclyl where R^(c) is an alkylene chain as defined above.If the heterocyclyl is a nitrogen-containing heterocyclyl, theheterocyclyl is attached to the alkyl radical at the nitrogen atom. Thealkylene chain of the heterocyclylalkyl radical is optionallysubstituted as defined above for an alkylene chain. The heterocyclylpart of the heterocyclylalkyl radical is optionally substituted asdefined above for a heterocyclyl group.

“Heteroaryl” refers to a radical derived from a 3- to 18-memberedaromatic ring radical that comprises two to seventeen carbon atoms andfrom one to six heteroatoms selected from nitrogen, oxygen and sulfur.As used herein, the heteroaryl radical is a monocyclic, bicyclic,tricyclic or tetracyclic ring system, wherein at least one of the ringsin the ring system is fully unsaturated, i.e., it contains a cyclic,delocalized (4n+2) π-electron system in accordance with the Hückeltheory. Heteroaryl groups include fused or bridged ring systems. Theheteroatoms in the heteroaryl radical are optionally oxidized. One ormore nitrogen atoms, if present, are optionally quaternized. Theheteroaryl is attached to the rest of the molecule through any atom ofthe ring(s). Examples of heteroaryls include, but are not limited to,azepinyl, acridinyl, benzimidazolyl, benzindolyl, 1,3-benzodioxolyl,benzofuranyl, benzooxazolyl, benzo[d]thiazolyl, benzothiadiazolyl,benzo[b][1,4]dioxepinyl, benzo[b][1,4]oxazinyl, 1,4-benzodioxanyl,benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl,benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl(benzothiophenyl), benzothieno[3,2-d]pyrimidinyl, benzotriazolyl,benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl,cyclopenta[d]pyrimidinyl,6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl,5,6-dihydrobenzo[h]quinazolinyl, 5,6-dihydrobenzo[h]cinnolinyl,6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazinyl, dibenzofuranyl,dibenzothiophenyl, furanyl, furanonyl, furo[3,2-c]pyridinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyrimidinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyridazinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyridinyl, isothiazolyl, imidazolyl,indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl,isoquinolyl, indolizinyl, isoxazolyl,5,8-methano-5,6,7,8-tetrahydroquinazolinyl, naphthyridinyl,1,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl,5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1-phenyl-1H-pyrrolyl,phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl,purinyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyridinyl,pyrido[3,2-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrazinyl,pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl,quinolinyl, isoquinolinyl, tetrahydroquinolinyl,5,6,7,8-tetrahydroquinazolinyl,5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl,6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinyl,5,6,7,8-tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl, thiadiazolyl,triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl,thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pridinyl, and thiophenyl (i.e.thienyl). Unless stated otherwise specifically in the specification, theterm “heteroaryl” includes heteroaryl radicals as defined above whichare optionally substituted by one or more substituents selected fromalkyl, alkenyl, alkynyl, halo, fluoroalkyl, haloalkenyl, haloalkynyl,oxo, thioxo, cyano, nitro, optionally substituted aryl, optionallysubstituted aralkyl, optionally substituted aralkenyl, optionallysubstituted aralkynyl, optionally substituted carbocyclyl, optionallysubstituted carbocyclylalkyl, optionally substituted heterocyclyl,optionally substituted heterocyclylalkyl, optionally substitutedheteroaryl, optionally substituted heteroarylalkyl, R^(b) OR^(a), R^(b)SR^(a), —R^(b)—OC(O)—R^(a), —R^(b)—N(R^(a))₂, —R^(b)—C(O)R^(a),—R^(b)—C(O)OR^(a), —R^(b)—C(O)N(R^(a))₂, —R^(b)—O—R^(c)—C(O)N(R^(a))₂,—R^(b)—N(R^(a))C(O)OR^(a), —R^(b)—N(R^(a))C(O)R^(a),—R^(b)—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —R^(b)—S(O)_(t)OR^(a)(where t is 1 or 2) and —R^(b)—S(O)_(t)N(R^(a))₂ (where t is 1 or 2),where each R^(a) is independently hydrogen, alkyl, fluoroalkyl,cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl,heterocyclylalkyl, heteroaryl or heteroarylalkyl, each R^(b) isindependently a direct bond or a straight or branched alkylene oralkenylene chain, and R^(c) is a straight or branched alkylene oralkenylene chain, and where each of the above substituents isunsubstituted unless otherwise indicated.

“N-heteroaryl” refers to a heteroaryl radical as defined abovecontaining at least one nitrogen and where the point of attachment ofthe heteroaryl radical to the rest of the molecule is through a nitrogenatom in the heteroaryl radical. An N-heteroaryl radical is optionallysubstituted as described above for heteroaryl radicals.

“Heteroarylalkyl” refers to a radical of the formula —R^(c)-heteroaryl,where R^(c) is an alkylene chain as defined above. If the heteroaryl isa nitrogen-containing heteroaryl, the heteroaryl is attached to thealkyl radical at the nitrogen atom. The alkylene chain of theheteroarylalkyl radical is optionally substituted as defined above foran alkylene chain. The heteroaryl part of the heteroarylalkyl radical isoptionally substituted as defined above for a heteroaryl group.

The compounds, or their pharmaceutically acceptable salts may containone or more asymmetric centers and may thus give rise to enantiomers,diastereomers, and other stereoisomeric forms that may be defined, interms of absolute stereochemistry, as (R)- or (S)- or, as (D)- or (L)-for amino acids. When the compounds described herein contain olefinicdouble bonds or other centers of geometric asymmetry, and unlessspecified otherwise, it is intended that the compounds include both Eand Z geometric isomers (e.g., cis or trans.) Likewise, all possibleisomers, as well as their racemic and optically pure forms, and alltautomeric forms are also intended to be included.

A “stereoisomer” refers to a compound made up of the same atoms bondedby the same bonds but having different three-dimensional structures,which are not interchangeable. It is therefore contemplated that variousstereoisomers and mixtures thereof and includes “enantiomers,” whichrefers to two stereoisomers whose molecules are nonsuperimposable mirrorimages of one another.

A “tautomer” refers to a proton shift from one atom of a molecule toanother atom of the same molecule. The compounds presented herein mayexist as tautomers. Tautomers are compounds that are interconvertible bymigration of a hydrogen atom, accompanied by a switch of a single bondand adjacent double bond. In solutions where tautomerization ispossible, a chemical equilibrium of the tautomers will exist. The exactratio of the tautomers depends on several factors, includingtemperature, solvent, and pH. Some examples of tautomeric pairs include:

“Optional” or “optionally” means that a subsequently described event orcircumstance may or may not occur and that the description includesinstances when the event or circumstance occurs and instances in whichit does not. For example, “optionally substituted aryl” means that thearyl radical may or may not be substituted and that the descriptionincludes both substituted aryl radicals and aryl radicals having nosubstitution.

“Pharmaceutically acceptable salt” includes both acid and base additionsalts. A pharmaceutically acceptable salt of any one of the styrenylderivative compounds described herein is intended to encompass any andall pharmaceutically suitable salt forms. Preferred pharmaceuticallyacceptable salts of the compounds described herein are pharmaceuticallyacceptable acid addition salts and pharmaceutically acceptable baseaddition salts.

“Pharmaceutically acceptable acid addition salt” refers to those saltswhich retain the biological effectiveness and properties of the freebases, which are not biologically or otherwise undesirable, and whichare formed with inorganic acids such as hydrochloric acid, hydrobromicacid, sulfuric acid, nitric acid, phosphoric acid, hydroiodic acid,hydrofluoric acid, phosphorous acid, and the like. Also included aresalts that are formed with organic acids such as aliphatic mono- anddicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoicacids, alkanedioic acids, aromatic acids, aliphatic and. aromaticsulfonic acids, etc. and include, for example, acetic acid,trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid,oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid,tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid,salicylic acid, and the like. Exemplary salts thus include sulfates,pyrosulfates, bisulfates, sulfites, bisulfites, nitrates, phosphates,monohydrogenphosphates, dihydrogenphosphates, metaphosphates,pyrophosphates, chlorides, bromides, iodides, acetates,trifluoroacetates, propionates, caprylates, isobutyrates, oxalates,malonates, succinate suberates, sebacates, fumarates, maleates,mandelates, benzoates, chlorobenzoates, methylbenzoates,dinitrobenzoates, phthalates, benzenesulfonates, toluenesulfonates,phenylacetates, citrates, lactates, malates, tartrates,methanesulfonates, and the like. Also contemplated are salts of aminoacids, such as arginates, gluconates, and galacturonates (see, forexample, Berge S. M. et al., “Pharmaceutical Salts,” Journal ofPharmaceutical Science, 66:1-19 (1997), which is hereby incorporated byreference in its entirety). Acid addition salts of basic compounds maybe prepared by contacting the free base forms with a sufficient amountof the desired acid to produce the salt according to methods andtechniques with which a skilled artisan is familiar.

“Pharmaceutically acceptable base addition salt” refers to those saltsthat retain the biological effectiveness and properties of the freeacids, which are not biologically or otherwise undesirable. These saltsare prepared from addition of an inorganic base or an organic base tothe free acid. Pharmaceutically acceptable base addition salts may beformed with metals or amines, such as alkali and alkaline earth metalsor organic amines. Salts derived from inorganic bases include, but arenot limited to, sodium, potassium, lithium, ammonium, calcium,magnesium, iron, zinc, copper, manganese, aluminum salts and the like.Salts derived from organic bases include, but are not limited to, saltsof primary, secondary, and tertiary amines, substituted amines includingnaturally occurring substituted amines, cyclic amines and basic ionexchange resins, for example, isopropylamine, trimethylamine,diethylamine, triethylamine, tripropylamine, ethanolamine,diethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol,dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine,N,N-dibenzylethylenediamine, chloroprocaine, hydrabamine, choline,betaine, ethylenediamine, ethylenedianiline, N-methylglucamine,glucosamine, methylglucamine, theobromine, purines, piperazine,piperidine, N-ethylpiperidine, polyamine resins and the like. See Bergeet al., supra.

“Non-retinoid compound” refers to any compound that is not a retinoid. Aretinoid is a compound that has a diterpene skeleton possessing atrimethylcyclohexenyl ring and a polyene chain that terminates in apolar end group. Examples of retinoids include retinaldehyde and derivedimine/hydrazide/oxime, retinol and any derived ester, retinyl amine andany derived amide, retinoic acid and any derived ester or amide. Anon-retinoid compound can comprise though not require an internal cyclicgroup (e.g., aromatic group). A non-retinoid compound can contain thoughnot require a styrenyl group.

“Prodrugs” is meant to indicate a compound that may be converted underphysiological conditions or by solvolysis to a biologically activecompound described herein. Thus, the term “prodrug” refers to aprecursor of a biologically active compound that is pharmaceuticallyacceptable. A prodrug may be inactive when administered to a subject,but is converted in vivo to an active compound, for example, byhydrolysis. The prodrug compound often offers advantages of solubility,tissue compatibility or delayed release in a mammalian organism (see,e.g., Bundgard, H., Design of Prodrugs (1985), pp. 7-9, 21-24 (Elsevier,Amsterdam).

A discussion of prodrugs is provided in Higuchi, T., et al., “Pro-drugsas Novel Delivery Systems,” A.C.S. Symposium Series, Vol. 14, and inBioreversible Carriers in Drug Design, ed. Edward B. Roche, AmericanPharmaceutical Association and Pergamon Press, 1987, both of which areincorporated in full by reference herein.

As used herein, “treatment” or “treating,” or “palliating” or“ameliorating” are used interchangeably herein. These terms refers to anapproach for obtaining beneficial or desired results including but notlimited to therapeutic benefit and/or a prophylactic benefit. Bytherapeutic benefit is meant eradication or amelioration of theunderlying disorder being treated. Also, a therapeutic benefit isachieved with the eradication or amelioration of one or more of thephysiological symptoms associated with the underlying disorder such thatan improvement is observed in the patient, notwithstanding that thepatient may still be afflicted with the underlying disorder. Forprophylactic benefit, the compositions may be administered to a patientat risk of developing a particular disease, or to a patient reportingone or more of the physiological symptoms of a disease, even though adiagnosis of this disease may not have been made.

The term “prodrug” is also meant to include any covalently bondedcarriers, which release the active compound in vivo when such prodrug isadministered to a mammalian subject. Prodrugs of an active compound, asdescribed herein, may be prepared by modifying functional groups presentin the active compound in such a way that the modifications are cleaved,either in routine manipulation or in vivo, to the parent activecompound. Prodrugs include compounds wherein a hydroxy, amino ormercapto group is bonded to any group that, when the prodrug of theactive compound is administered to a mammalian subject, cleaves to forma free hydroxy, free amino or free mercapto group, respectively.Examples of prodrugs include, but are not limited to, acetate, formateand benzoate derivatives of alcohol or amine functional groups in theactive compounds and the like.

Preparation of the Styrenyl Derivative Compounds

In general, the compounds used in the reactions described herein aremade according to organic synthesis techniques known to those skilled inthis art, starting from commercially available chemicals and/or fromcompounds described in the chemical literature. “Commercially availablechemicals” are obtained from standard commercial sources including AcrosOrganics (Pittsburgh Pa.), Aldrich Chemical (Milwaukee Wis., includingSigma Chemical and Fluka), Apin Chemicals Ltd. (Milton Park UK), AvocadoResearch (Lancashire U.K.), BDH Inc. (Toronto, Canada), Bionet(Cornwall, U.K.), Chemservice Inc. (West Chester Pa.), Crescent ChemicalCo. (Hauppauge N.Y.), Eastman Organic Chemicals, Eastman Kodak Company(Rochester N.Y.), Fisher Scientific Co. (Pittsburgh Pa.), FisonsChemicals (Leicestershire UK), Frontier Scientific (Logan Utah), ICNBiomedicals, Inc. (Costa Mesa Calif.), Key Organics (Cornwall U.K.),Lancaster Synthesis (Windham N.H.), Maybridge Chemical Co. Ltd.(Cornwall U.K.), Parish Chemical Co. (Orem Utah), Pfaltz & Bauer, Inc.(Waterbury Conn.), Polyorganix (Houston Tex.), Pierce Chemical Co.(Rockford Ill.), Riedel de Haen A G (Hanover, Germany), Spectrum QualityProduct, Inc. (New Brunswick, N.J.), TCI America (Portland Oreg.), TransWorld Chemicals, Inc. (Rockville Md.), and Wako Chemicals USA, Inc.(Richmond Va.).

Methods known to one of ordinary skill in the art are identified throughvarious reference books and databases. Suitable reference books andtreatise that detail the synthesis of reactants useful in thepreparation of compounds described herein, or provide references toarticles that describe the preparation, include for example, “SyntheticOrganic Chemistry”, John Wiley & Sons, Inc., New York; S. R. Sandler etal., “Organic Functional Group Preparations,” 2nd Ed., Academic Press,New York, 1983; H. O. House, “Modern Synthetic Reactions”, 2nd Ed., W.A. Benjamin, Inc. Menlo Park, Calif. 1972; T. L. Gilchrist,“Heterocyclic Chemistry”, 2nd Ed., John Wiley & Sons, New York, 1992; J.March, “Advanced Organic Chemistry: Reactions, Mechanisms andStructure”, 4th Ed., Wiley-Interscience, New York, 1992. Additionalsuitable reference books and treatise that detail the synthesis ofreactants useful in the preparation of compounds described herein, orprovide references to articles that describe the preparation, includefor example, Fuhrhop, J. and Penzlin G “Organic Synthesis: Concepts,Methods, Starting Materials”, Second, Revised and Enlarged Edition(1994) John Wiley & Sons ISBN: 3-527-29074-5; Hoffman, R. V. “OrganicChemistry, An Intermediate Text” (1996) Oxford University Press, ISBN0-19-509618-5; Larock, R. C. “Comprehensive Organic Transformations: AGuide to Functional Group Preparations” 2nd Edition (1999) Wiley-VCH,ISBN: 0-471-19031-4; March, J. “Advanced Organic Chemistry: Reactions,Mechanisms, and Structure” 4th Edition (1992) John Wiley & Sons, ISBN:0-471-60180-2; Otera, J. (editor) “Modern Carbonyl Chemistry” (2000)Wiley-VCH, ISBN: 3-527-29871-1; Patai, S. “Patai's 1992 Guide to theChemistry of Functional Groups” (1992) Interscience ISBN: 0-471-93022-9;Quin, L. D. et al. “A Guide to Organophosphorus Chemistry” (2000)Wiley-Interscience, ISBN: 0-471-31824-8; Solomons, T. W. G. “OrganicChemistry” 7th Edition (2000) John Wiley & Sons, ISBN: 0-471-19095-0;Stowell, J. C., “Intermediate Organic Chemistry” 2nd Edition (1993)Wiley-Interscience, ISBN: 0-471-57456-2; “Industrial Organic Chemicals:Starting Materials and Intermediates: An Ullmann's Encyclopedia” (1999)John Wiley & Sons, ISBN: 3-527-29645-X, in 8 volumes; “OrganicReactions” (1942-2000) John Wiley & Sons, in over 55 volumes; and“Chemistry of Functional Groups” John Wiley & Sons, in 73 volumes.

Specific and analogous reactants may also be identified through theindices of known chemicals prepared by the Chemical Abstract Service ofthe American Chemical Society, which are available in most public anduniversity libraries, as well as through on-line databases (the AmericanChemical Society, Washington, D.C., may be contacted for more details).Chemicals that are known but not commercially available in catalogs maybe prepared by custom chemical synthesis houses, where many of thestandard chemical supply houses (e.g., those listed above) providecustom synthesis services. A reference for the preparation and selectionof pharmaceutical salts of the styrenyl derivative compounds describedherein is P. H. Stahl & C. G. Wermuth “Handbook of PharmaceuticalSalts”, Verlag Helvetica Chimica Acta, Zurich, 2002.

Generally speaking, the compounds disclosed herein can be prepared in astepwise manner involving an olefin formation and a side chainformation.

In certain embodiments, an olefin intermediate can be first constructed,which forms the precursor to the styrenyl core structure. A side chainmoiety, which is a precursor to the linkage and the nitrogen-containingmoiety of the compounds disclosed herein, can then be attached to theolefin intermediate.

In other embodiments, the compounds disclosed herein can be prepared byfirst preparing a phenyl intermediate having an appropriate side chain,followed by an olefin formation to provide the styrenyl core structure.

The following Methods illustrate various synthetic pathways forpreparing olefin intermediates and the side chain moieties. One skilledin the art will recognize that a method for olefin formation can becombined with a method for side chain formation to provide the compoundsdisclosed herein. For example, Method A can be combined with any ofMethod K, Methods K and U, Methods K and L, Methods K and AB, Methods Tand L, Method R, Method S, Method J, Method E, Methods R and U, and thelike. Similarly, Method C can be combined with Method J.

Olefin Formation:

Methods A-I below describe various approaches to olefin formation.

More specifically, Method A illustrates constructing an olefinintermediate (A-3) in a Wittig reaction. Depending on the sequence ofthe reactions, Ar can be a phenyl derivative compound that is alreadyattached to a side chain moiety, or Ar may comprise a reactive group(appropriately protected), which will be coupled to a side chain moietyafter the olefin formation step.

According to Method A, a phosphonium ylide reagent (or “Wittig reagent”)(A-1) can be coupled to a benzaldehyde or ketone derivative (A-2) toprovide the olefin intermediate (A-3) in the presence of a base. Thegeometry of the resulting A-3 may depend on the reactivity of the ylidereagent. Triphenylphosphonium-based ylide reagent (R is phenyl)typically produces predominantly (E) or trans-styrenes; whereastrialkylphosphonium-based ylide reagent (R is alkyl) producespredominantly (Z) or cis-styrene. The E or Z stereoisomers can beseparated by, for example, chromatography or other known methods in theart.

The ylide reagent (A-1) can be prepared according to known methods inthe art. For example, R₁₁—CH₂OH can be converted to the correspondingylide reagent (A-1) in the presence of triphenylphosphine hydrobromide.The benzaldehyde or ketone derivative (A-2) may be commerciallyavailable or can be prepared by known methods in the art.

The olefin intermediate (A-3) may also be prepared by coupling aphosphonium ylide reagent derivatized from the Ar group (A-4) and analdehyde or ketene derivative of R₁₁ (A-5). The ylide reagent (A-4) canbe prepared from, for example, a benzyl alcohol, whereas (A-5) can beprepared by known methods in the art or can be obtained from commercialvendors.

Method AE shows a coupling reaction similar to the Wittig reaction ofMethod A, except that a phosphorus ylide is used in place of thephosphonium ylide. The phosphorus ylide can be coupled to an aldehyde orketone in the presence of a base (Wittig-Horner-Emmons reaction.)

In addition, elimination reactions can be used to form olefin bonds.Methods B-D illustrate various approaches to forming alcohol precursorsthat can undergo alcohol dehydration in acidic conditions to produceolefin bonds. The Ar group is typically activated with a metal (e.g.,Li) to facilitate the alcohol formation. Grignard reagent can also beused in place of the metal.

As discussed above in connection with Method A, the alcohol precursor ineach of Methods B-D can also be prepared by using a metal activated R₁₁group and an Ar group derivatized with a carbonyl group or a cyclopropylgroup.

Methods E-G illustrate coupling an olefin or an activated olefindirectly with an aryl halide in the presence of a palladium(0) catalyst.In certain embodiments, the olefin can be activated by a transitionmetal (e.g., Zn or Sn), or boronic acid (e.g., Suzuki reaction), as areknown in the art. The halo substituent of the aryl group can be, forexample, bromo or iodo.

Palladium catalysts suitable for coupling reactions are known to oneskilled in the art. Exemplary palladium(0) catalysts include, forexample, tetrakis(triphenylphosphine)palladium(0) [Pd(PPh₃)₄] andtetrakis(tri(o-tolylphosphine)palladium(0),tetrakis(dimethylphenylphosphine)palladium(0),tetrakis(tris-p-methoxyphenylphosphine)palladium(0) and the like. It isunderstood that a palladium (II) salt can also be used, which generatesthe palladium (0) catalyst in situ. Suitable palladium (II) saltsinclude, for example, palladium diacetate [Pd(OAc)₂],bis(triphenylphosphine)-palladium diacetate and the like.

An olefin intermediate can also be constructed from an alkyneaddition/hydrogenation reaction. Depending on the reaction conditions(syn or anti addition), cis or trans configuration can be formed.

Method H illustrates a syn-addition, i.e., both hydrogens are added fromone side of the alkyne molecule, which results in a cis olefinconfiguration. Typically, hydrogen gas can be used in the presence of acatalyst (e.g., Pd on carbon or platinum) to effect a syn addition.

Method I illustrate an anti-addition, i.e., an adding agent is added toopposite sides of the alkyne molecule, resulting in a trans olefinconfiguration. The adding agent can be, for example, aluminum hydridereagents, lithium/NH₃ reagents and the like.

Side Chain Formation and Modification

Methods J-T and AA-AD below describe various approaches to side chainformation and modifications.

Generally speaking, a suitably substituted phenyl derivative can becoupled to a diverse range of side chains, which may be further modifiedto provide the final linkages and the nitrogen-containing moieties ofthe compounds disclosed herein.

Method J illustrates an aryl halide coupled with an allyl alcohol in thepresence of a palladium(0) catalyst. The terminal alcohol group of allylalcohol has been simultaneously oxidized to an aldehyde group, which canbe further reduced to an amine (—NR₉R₁₀).

Method K illustrates an aldol condensation between an aryl aldehyde oraryl ketone with a nitrile reagent comprising at least one α-hydrogen.The resulting condensation intermediate can be further reduced to anamine (—NR₉R₁₀).

Method AA shows an acylation reaction to form a ketone-based linkage.One skilled in the art will recognize that the R′ group may comprisefunctional groups that can be further modified.

Method R shows a ring-opening reaction of an epoxide reagent to form a3-carbon side chain linkage.

Method S shows the formation of a triple bond linkage based on aSonogashira reaction. Typically, palladium(0) catalyst is used incombination with a base to couple an aryl halide with a acetylenederivative. R′ can be further modified, as described herein.

Method T shows the formation of a double bond linkage based on a Heckreaction. Typically, palladium(0) catalyst is used in combination with abase to couple an aryl halide with a vinyl derivative. R′ can be furthermodified, as described herein.

Methods M-P illustrate attachments of side chain moieties byheteroatoms. Method M shows a side chain precursor (R′OH) attached to anaryl derivative via an oxygen atom in a condensation reaction in which amolecule of H₂O is eliminated. R′ may comprise functional groups thatcan be further modified to prepare linkages and nitrogen-containingmoieties of the compounds disclosed herein.

Method N shows a similar coupling reaction that provides a sulfurlinking atom. Method O illustrates an oxidation step of the sulfurlinking atom to provide —S(O)— or —S(O)₂—, depending on the degree ofoxidation.

Method P shows the formation of an amide-containing linkage, in which aaniline derivative is coupled with a carboxylic acid derivative. Thecarboxylic acid derivative can be activated to facilitate the amideformation. Suitable activating reagents include, for example,1,3-dicyclohexylcarbodiimide (DCC), 1,1′-carbonyldiimidazole (CDI),1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCL),benzotriazol-1-yl-oxy-tris(dimethylamino)phosphonium hexafluorophosphate(BOP), and 1,3-diisopropylcarbodiimide (DICD).

After attachment, the side chain moiety can be further modified toprovide the final linkage and the terminal nitrogen-containing moietyfor the compounds disclosed herein. The following methods illustrate avariety of synthetic pathways to manipulate or modify the side chainmoiety by reduction, oxidation, nucleophilic or electrophilicsubstitution, fluorination, acylation and the like. As a result, adiverse group of linkages can be synthesized.

Method L illustrates an amination process in which carboxylic acid isconverted to an amine. Typically, the carboxylic acid (or ester) can befirst reduced to primary alcohol, which can then be converted to anamine via mesylate, halide, azide, phthalimide, or Mitsunobu reactionand the like. Suitable reducing agents include, for example, sodiumborohydride (NaBH₄), sodium cyanoborohydride (NaBH₃CN), sodiumtriacetoxyborohydride (NaBH(OCOCH₃)₃), lithium aluminum hydride (LiAlH₄)and the like. As shown, the resulting amine can be furtherfunctionalized, by known methods in the art.

Additional or alternative modifications can be carried out according tothe methods illustrated below.

Scheme I illustrates a complete synthetic sequence for preparing oneexample of the compounds disclosed herein.

In Scheme I, an olefin intermediate is first constructed, followed bycoupling to a side chain moiety. Further modification of the side chainmoiety by reduction affords the compounds disclosed herein having apropylene linkage and a terminal amine. Other nitrogen-containingmoieties can be further derived from the terminal amine, according toknown methods in the art.

One skilled in the art should recognize, however, that the order of thereactions may vary. Thus, in other embodiments, as shown in Scheme II, aside chain attachment is initially performed, followed by olefinformation.

In addition to the generic reaction schemes and methods discussed above,other exemplary reaction schemes are also provided to illustrate methodsfor preparing any of the compounds disclosed herein.

Treatment of Ophthalmic Diseases and Other Disorders

Styrenyl derivative compounds having a structure of the specificcompounds described herein may be useful for treating an ophthalmicdisease or other disorder. One or more of the subject compounds mayinhibit one or more steps in the visual cycle, for example, byinhibiting or blocking a functional activity of a visual cycle trans-cisisomerase (also including a visual cycle trans-cis isomerohydrolase).The compounds described herein, may inhibit, block, or in some mannerinterfere with the isomerization step in the visual cycle. In aparticular embodiment, the compound inhibits isomerization of anall-trans-retinyl ester; in certain embodiments, the all-trans-retinylester is a fatty acid ester of all-trans-retinol, and the compoundinhibits isomerization of all-trans-retinol to 11-cis-retinol. Thecompound may bind to, or in some manner interact with, and inhibit theisomerase activity of at least one visual cycle isomerase, which mayalso be referred to herein and in the art as a retinal isomerase or anisomerohydrolase. The compound may block or inhibit binding of anall-trans-retinyl ester substrate to an isomerase. Alternatively, or inaddition, the compound may bind to the catalytic site or region of theisomerase, thereby inhibiting the capability of the enzyme to catalyzeisomerization of an all-trans-retinyl ester substrate. In general, atleast one isomerase that catalyzes the isomerization ofall-trans-retinyl esters is believed to be located in the cytoplasm ofRPE cells.

A method for determining the effect of a compound on isomerizationprocess may be performed in vitro as described herein and in the art(Stecher et al., J Biol Chem 274:8577-85 (1999); see also Golczak etal., Proc. Natl. Acad. Sci. USA 102:8162-67 (2005)). Retinal pigmentepithelium (RPE) microsome membranes isolated from an animal (such asbovine, porcine, human, for example) may serve as the source of theisomerase. The capability of the styrenyl derivative compounds toinhibit isomerase may also be determined by an in vivo murine isomeraseassay. Brief exposure of the eye to intense light (“photobleaching” ofthe visual pigment or simply “bleaching”) is known to photo-isomerizealmost all 11-cis-retinal in the retina. The recovery of 11-cis-retinalafter bleaching can be used to estimate the activity of isomerase invivo (see, e.g., Maeda et al., J. Neurochem 85:944-956 (2003); VanHooser et al., J Biol Chem 277:19173-82, 2002). Electroretinographic(ERG) recording may be performed as previously described (Haeseleer etal., Nat. Neurosci. 7:1079-87 (2004); Sugitomo et al., J. Toxicol. Sci.22 Suppl 2:315-25 (1997); Keating et al., Documenta Ophthalmologica100:77-92 (2000)). See also Deigner et al., Science, 244: 968-971(1989); Gollapalli et al., Biochim Biophys Acta. 1651: 93-101 (2003);Parish, et al., Proc. Natl. Acad. Sci. USA 95:14609-13 (1998); Radu, etal., Proc Natl Acad Sci USA 101: 5928-33 (2004)). In certainembodiments, compounds that are useful for treating a subject who has orwho is at risk of developing any one of the ophthalmic and retinaldiseases or disorders described herein have IC₅₀ levels (compoundconcentration at which 50% of isomerase activity is inhibited) asmeasured in the isomerase assays described herein or known in the artthat is less than about 1 μM; in other embodiments, the determined IC₅₀level is less than about 10 nM; in other embodiments, the determinedIC₅₀ level is less than about 50 nM; in certain other embodiments, thedetermined IC₅₀ level is less than about 100 nM; in other certainembodiments, the determined IC₅₀ level is less than about 10 μM; inother embodiments, the determined IC₅₀ level is less than about 50 μM;in other certain embodiments, the determined IC₅₀ level is less thanabout 100 μM or about 500 μM; in other embodiments, the determined IC₅₀level is between about 1 μM and 10 μM; in other embodiments, thedetermined IC₅₀ level is between about 1 nM and 10 nM. When adminsteredinto a subject, one or more compounds of the present invention exhibitsan ED50 value of about 5 mg/kg, 5 mg/kg or less as ascertained byinhibition of an isomerase reaction that results in production of 11-cisretinol. In some embodiments, the compounds of the present inventionhave ED50 values of about 1 mg/kg when administered into a subject. Inother embodiments, the compounds of the present invention have ED50values of about 0.1 mg/kg when administered into a subject. The ED50values can be measured after about 2 hours, 4 hours, 6 hours, 8 hours orlonger upon administering a subject compound or a pharmaceuticalcomposition thereof. The compounds described herein may be useful fortreating a subject who has an ophthalmic disease or disorder such asage-related macular degeneration or Stargardt's macular dystrophy. Inone embodiment, the compounds described herein may inhibit (i.e.,prevent, reduce, slow, abrogate, or minimize) accumulation of lipofuscinpigments and lipofuscin-related and/or associated molecules in the eye.In another embodiment, the compounds may inhibit (i.e., prevent, reduce,slow, abrogate, or minimize) N-retinylidene-N-retinylethanolamine (A2E)accumulation in the eye. The ophthalmic disease may result, at least inpart, from lipofuscin pigments accumulation and/or from accumulation ofA2E in the eye. Accordingly, in certain embodiments, methods areprovided for inhibiting or preventing accumulation of lipofuscinpigments and/or A2E in the eye of a subject. These methods compriseadministering to the subject a composition comprising a pharmaceuticallyacceptable or suitable excipient (i.e., pharmaceutically acceptable orsuitable carrier) and a styrenyl derivative compound as described indetail herein.

Accumulation of lipofuscin pigments in retinal pigment epithelium (RPE)cells has been linked to progression of retinal diseases that result inblindness, including age-related macular degeneration (De Lacy et al.,Retina 15:399-406 (1995)). Lipofuscin granules are autofluorescentlysosomal residual bodies (also called age pigments). The majorfluorescent species of lipofuscin is A2E (an orange-emittingfluorophore), which is a positively charged Schiff-basecondensation-product formed by all-trans retinaldehyde withphosphatidylethanolamine (2:1 ratio) (see, e.g., Eldred et al., Nature361:724-6 (1993); see also, Sparrow, Proc. Natl. Acad. Sci. USA100:4353-54 (2003)). Much of the indigestible lipofuscin pigment isbelieved to originate in photoreceptor cells; deposition in the RPEoccurs because the RPE internalize membranous debris that is discardeddaily by the photoreceptor cells. Formation of this compound is notbelieved to occur by catalysis by any enzyme, but rather A2E forms by aspontaneous cyclization reaction. In addition, A2E has a pyridiniumbisretinoid structure that once formed may not be enzymaticallydegraded. Lipofuscin, and thus A2E, accumulate with aging of the humaneye and also accumulate in a juvenile form of macular degenerationcalled Stargardt's disease, and in several other congenital retinaldystrophies.

A2E may induce damage to the retina via several different mechanisms. Atlow concentrations, A2E inhibits normal proteolysis in lysosomes (Holzet al., Invest. Ophthalmol. Vis. Sci. 40:737-43 (1999)). At higher,sufficient concentrations, A2E may act as a positively chargedlysosomotropic detergent, dissolving cellular membranes, and may alterlysosomal function, release proapoptotic proteins from mitochondria, andultimately kill the RPE cell (see, e.g., Eldred et al., supra; Sparrowet al., Invest. Ophthalmol. Vis. Sci. 40:2988-95 (1999); Holz et al.,supra; Finneman et al., Proc. Natl. Acad. Sci. USA 99:3842-347 (2002);Suter et al., J. Biol. Chem. 275:39625-30 (2000)). A2E is phototoxic andinitiates blue light-induced apoptosis in RPE cells (see, e.g., Sparrowet al., Invest. Ophthalmol. Vis. Sci. 43:1222-27 (2002)). Upon exposureto blue light, photooxidative products of A2E are formed (e.g.,epoxides) that damage cellular macromolecules, including DNA (Sparrow etal., J. Biol. Chem. 278(20):18207-13 (2003)). A2E self-generates singletoxygen that reacts with A2E to generate epoxides at carbon-carbon doublebonds (Sparrow et al., supra). Generation of oxygen reactive speciesupon photoexcitation of A2E causes oxidative damage to the cell, oftenresulting in cell death. An indirect method of blocking formation of A2Eby inhibiting biosynthesis of the direct precursor of A2E,all-trans-retinal, has been described (see U.S. Patent ApplicationPublication No. 2003/0032078). However, the usefulness of the methoddescribed therein is limited because generation of all-trans retinal isan important component of the visual cycle. Other therapies describedinclude neutralizing damage caused by oxidative radical species by usingsuperoxide-dismutase mimetics (see, e.g., U.S. Patent ApplicationPublication No. 2004/0116403) and inhibiting A2E-induced cytochrome Coxidase in retinal cells with negatively charged phospholipids (see,e.g., U.S. Patent Application Publication No. 2003/0050283).

The styrenyl derivative compounds described herein may be useful forpreventing, reducing, inhibiting, or decreasing accumulation (i.e.,deposition) of A2E and A2E-related and/or derived molecules in the RPE.Without wishing to be bound by theory, because the RPE is critical forthe maintenance of the integrity of photoreceptor cells, preventing,reducing, or inhibiting damage to the RPE may inhibit degeneration(enhance the survival or increase cell viability) of retinal neuronalcells, particularly, photoreceptor cells. Compounds that bindspecifically to or interact with A2E A2E-related and/or derivedmolecules or that affect A2E formation or accumulation may also reduce,inhibit, prevent, or decrease one or more toxic effects of A2E or ofA2E-related and/or derived molecules that result in retinal neuronalcell (including a photoreceptor cell) damage, loss, orneurodegeneration, or in some manner decrease retinal neuronal cellviability. Such toxic effects include induction of apoptosis,self-generation of singlet oxygen and generation of oxygen reactivespecies; self-generation of singlet oxygen to form A2E-epoxides thatinduce DNA lesions, thus damaging cellular DNA and inducing cellulardamage; dissolving cellular membranes; altering lysosomal function; andeffecting release of proapoptotic proteins from mitochondria.

In other embodiments, the compounds described herein may be used fortreating other ophthalmic diseases or disorders, for example, glaucoma,retinal detachment, cone-rod dystrophy, hemorrhagic or hypertensiveretinopathy, retinitis pigmentosa, optic neuropathy, inflammatoryretinal disease, proliferative vitreoretinopathy, genetic retinaldystrophies, traumatic injury to the optic nerve (such as by physicalinjury, excessive light exposure, or laser light), hereditary opticneuropathy, neuropathy due to a toxic agent or caused by adverse drugreactions or vitamin deficiency, Sorsby's fundus dystrophy, uveitis, aretinal disorder associated with Alzheimer's disease, a retinal disorderassociated with multiple sclerosis; a retinal disorder associated withviral infection (cytomegalovirus or herpes simplex virus), a retinaldisorder associated with Parkinson's disease, a retinal disorderassociated with AIDS, or other forms of progressive retinal atrophy ordegeneration. In another specific embodiment, the disease or disorderresults from mechanical injury, chemical or drug-induced injury, thermalinjury, radiation injury, light injury, laser injury. The subjectcompounds are useful for treating both hereditary and non-hereditaryretinal dystrophy. These methods are also useful for preventingophthalmic injury from environmental factors such as light-inducedoxidative retinal damage, laser-induced retinal damage, “flash bombinjury,” or “light dazzle,” refractive errors including but not limitedto myopia (see, e.g., Quinn G E et al. Nature 1999; 399:113-114; ZadnikK et al. Nature 2000; 404:143-144; Gwiazda J et al. Nature 2000; 404:144).

In other embodiments, methods are provided herein for inhibitingneovascularization (including but not limited to neovascular glaucoma)in the retina using any one or more of the styrenyl derivative compoundsdescribed herein, substructures thereof, and the specific styrenylcompounds recited herein). In certain other embodiments, methods areprovided for reducing hypoxia in the retina using the compoundsdescribed herein. These methods comprise administering to a subject, inneed thereof, a composition comprising a pharmaceutically acceptable orsuitable excipient (i.e., pharmaceutically acceptable or suitablecarrier) and a styrenyl derivative compound as described in detailherein, including a compound having the structure as set forth herein,substructures thereof, and the specific styrenyl compounds recitedherein.

Merely by way of explanation and without being bound by any theory, andas discussed in further detail herein, dark-adapted rod photoreceptorsengender a very high metabolic demand (i.e., expenditure of energy (ATPconsumption) and consumption of oxygen). The resultant hypoxia may causeand/or exacerbate retinal degeneration, which is likely exaggeratedunder conditions in which the retinal vasculature is alreadycompromised, including, but not limited to, such conditions as diabeticretinopathy, macular edema, diabetic maculopathy, retinal blood vesselocclusion (which includes retinal venous occlusion and retinal arterialocclusion), retinopathy of prematurity, ischemia reperfusion relatedretinal injury, as well as in the wet form of age-related maculardegeneration (AMD). Furthermore, retinal degeneration and hypoxia maylead to neovascularization, which in turn may worsen the extent ofretinal degeneration. The styrenyl derivative compounds described hereinthat modulate the visual cycle can be administered to prevent, inhibit,and/or delay dark adaptation of rod photoreceptor cells, and maytherefore reduce metabolic demand, thereby reducing hypoxia andinhibiting neovascularization.

By way of background, oxygen is a critical metabolite for preservationof retinal function in mammals, and retinal hypoxia may be a factor inmany retinal diseases and disorders that have ischemia as a component.In most mammals (including humans) with dual vascular supply to theretina, oxygenation of the inner retina is achieved through theintraretinal microvasculature, which is sparse compared to thechoriocapillaris that supplies oxygen to the RPE and photoreceptors. Thedifferent vascular supply networks create an uneven oxygen tensionacross the thickness of the retina (Cringle et al., Invest. Ophthalmol.Vis. Sci. 43:1922-27 (2002)). Oxygen fluctuation across the retinallayers is related to both the differing capillary densities anddisparity in oxygen consumption by various retinal neurons and glia.

Local oxygen tension can significantly affect the retina and itsmicrovasculature by regulation of an array of vasoactive agents,including, for example, vascular endothelial growth factor (VEGF). (See,e.g., Werdich et al., Exp. Eye Res. 79:623 (2004); Arden et al., Br. J.Ophthalmol. 89:764 (2005)). Rod photoreceptors are believed to have thehighest metabolic rate of any cell in the body (see, e.g., Arden et al.,supra). During dark adaptation, the rod photoreceptors recover theirhigh cytoplasmic calcium levels via cGMP-gated calcium channels withconcomitant extrusion of sodium ions and water. The efflux of sodiumfrom the cell is an ATP-dependent process, such that the retinal neuronsconsume up to an estimated five times more oxygen under scotopic (i.e.,dark adapted), compared with photopic (i.e., light adapted) conditions.Thus, during characteristic dark adaptation of photoreceptors, the highmetabolic demand leads to significant local reduction of oxygen levelsin the dark-adapted retina (Ahmed et al, Invest. Ophthalmol. Vis. Sci.34:516 (1993)).

Without being bound by any one theory, retinal hypoxia may be furtherincreased in the retina of subjects who have diseases or conditions suchas, for example, central retinal vein occlusion in which the retinalvasculature is already compromised. Increasing hypoxia may increasesusceptibility to sight-threatening, retinal neovascularization.Neovascularization is the formation of new, functional microvascularnetworks with red blood cell perfusion, and is a characteristic ofretinal degenerative disorders, including, but not limited to, diabeticretinopathy, retinopathy of prematurity, wet AMD and central retinalvein occlusions. Preventing or inhibiting dark adaptation of rodphotoreceptor cells, thereby decreasing expenditure of energy andconsumption of oxygen (i.e., reducing metabolic demand), may inhibit orslow retinal degeneration, and/or may promote regeneration of retinalcells, including rod photoreceptor cells and retinal pigment epithelial(RPE) cells, and may reduce hypoxia and may inhibit neovascularization.

Methods are described herein for inhibiting (i.e., reducing, preventing,slowing or retarding, in a biologically or statistically significantmanner) degeneration of retinal cells (including retinal neuronal cellsas described herein and RPE cells) and/or for reducing (i.e., preventingor slowing, inhibiting, abrogating in a biologically or statisticallysignificant manner) retinal ischemia. Methods are also provided forinhibiting (i.e., reducing, preventing, slowing or retarding, in abiologically or statistically significant manner) neovascularization inthe eye, particularly in the retina. Such methods comprise contactingthe retina, and thus, contacting retinal cells (including retinalneuronal cells such as rod photoreceptor cells, and RPE cells) with atleast one of the styrenyl derivative compounds described herein thatinhibits at least one visual cycle trans-cis isomerase (which mayinclude inhibition of isomerization of an all-trans-retinyl ester),under conditions and at a time that may prevent, inhibit, or delay darkadaptation of a rod photoreceptor cell in the retina. As described infurther detail herein, in particular embodiments, the compound thatcontacts the retina interacts with an isomerase enzyme or enzymaticcomplex in a RPE cell in the retina and inhibits, blocks, or in somemanner interferes with the catalytic activity of the isomerase. Thus,isomerization of an all-trans-retinyl ester is inhibited or reduced. Theat least one styrenyl derivative compound (or composition comprising atleast one compound) may be administered to a subject who has developedand manifested an ophthalmic disease or disorder or who is at risk ofdeveloping an ophthalmic disease or disorder, or to a subject whopresents or who is at risk of presenting a condition such as retinalneovascularization or retinal ischemia.

By way of background, the visual cycle (also called retinoid cycle)refers to the series of enzyme and light-mediated conversions betweenthe 11-cis and all-trans forms of retinol/retinal that occur in thephotoreceptor and retinal pigment epithelial (RPE) cells of the eye. Invertebrate photoreceptor cells, a photon causes isomerization of the11-cis-retinylidene chromophore to all-trans-retinylidene coupled to thevisual opsin receptors. This photoisomerization triggers conformationalchanges of opsins, which, in turn, initiate the biochemical chain ofreactions termed phototransduction (Filipek et al., Annu. Rev. Physiol.65 851-79 (2003)). After absorption of light and photoisomerization of11-cis-retinal to all-trans retinal, regeneration of the visualchromophore is a critical step in restoring photoreceptors to theirdark-adapted state. Regeneration of the visual pigment requires that thechromophore be converted back to the 11-cis-configuration (reviewed inMcBee et al., Prog. Retin Eye Res. 20:469-52 (2001)). The chromophore isreleased from the opsin and reduced in the photoreceptor by retinoldehydrogenases. The product, all-trans-retinol, is trapped in theadjacent retinal pigment epithelium (RPE) in the form of insoluble fattyacid esters in subcellular structures known as retinosomes (Imanishi etal., J. Cell Biol. 164:373-78 (2004)).

During the visual cycle in rod receptor cells, the 11-cis retinalchromophore within the visual pigment molecule, which is calledrhodopsin, absorbs a photon of light and is isomerized to the all-transconfiguration, thereby activating the phototransduction cascade.Rhodopsin is a G-protein coupled receptor (GPCR) that consists of sevenmembrane-spanning helices that are interconnected by extracellular andcytoplasmic loops. When the all-trans form of the retinoid is stillcovalently bound to the pigment molecule, the pigment is referred to asmetarhodopsin, which exists in different forms (e.g., metarhodopsin Iand metarhodopsin II). The all-trans retinoid is then hydrolyzed and thevisual pigment is in the form of the apoprotein, opsin, which is alsocalled apo-rhodopsin in the art and herein. This all-trans retinoid istransported or chaperoned out of the photoreceptor cell and across theextracellular space to the RPE cells, where the retinoid is converted tothe 11-cis isomer. The movement of the retinoids between the RPE andphotoreceptors cells is believed to be accomplished by differentchaperone polypeptides in each of the cell types. See Lamb et al.,Progress in Retinal and Eye Research 23:307-80 (2004).

Under light conditions, rhodopsin continually transitions through thethree forms, rhodopsin, metarhodopsin, and apo-rhodopsin. When most ofthe visual pigment is in the rhodopsin form (i.e., bound with 11-cisretinal), the rod photoreceptor cell is in a “dark-adapted” state. Whenthe visual pigment is predominantly in the metarhodopsin form (i.e.,bound with all-trans-retinal), the state of the photoreceptor cell isreferred to as a “light-adapted,” and when the visual pigment isapo-rhodopsin (or opsin) and no longer has bound chromophore, the stateof the photoreceptor cell is referred to as “rhodopsin-depleted.” Eachof the three states of the photoreceptor cell has different energyrequirements, and differing levels of ATP and oxygen are consumed. Inthe dark-adapted state, rhodopsin has no regulatory effect on cationchannels, which are open, resulting in an influx of cations (Na⁺/K⁺ andCa²⁺). To maintain the proper level of these cations in the cell duringthe dark state, the photoreceptor cells actively transport the cationsout of the cell via ATP-dependent pumps. Thus maintenance of this “darkcurrent” requires a large amount of energy, resulting in high metabolicdemand. In the light-adapted state, metarhodopsin triggers an enzymaticcascade process that results in hydrolysis of GMP, which in turn, closescation-specific channels in the photoreceptor cell membrane. In therhodopsin-depleted state, the chromophore is hydrolyzed frommetarhodopsin to form the apoprotein, opsin (apo-rhodopsin), whichpartially regulates the cation channels such that the rod photoreceptorcells exhibit an attenuated current compared with the photoreceptor inthe dark-adapted state, resulting in a moderate metabolic demand.

Under normal light conditions, the incidence of rod photoreceptors inthe dark adapted state is small, in general, 2% or less, and the cellsare primarily in the light-adapted or rhodopsin-depleted states, whichoverall results in a relatively low metabolic demand compared with cellsin the dark-adapted state. At night, however, the relative incidence ofthe dark-adapted photoreceptor state increases profoundly, due to theabsence of light adaptation and to the continued operation of the “dark”visual cycle in RPE cells, which replenishes the rod photoreceptor cellswith 11-cis-retinal. This shift to dark adaptation of the rodphotoreceptor causes an increase in metabolic demand (that is, increasedATP and oxygen consumption), leading ultimately to retinal hypoxia andsubsequent initiation of angiogenesis. Most ischaemic insults to theretina therefore occur in the dark, for example, at night during sleep.

Without being bound by any theory, therapeutic intervention during the“dark” visual cycle may prevent retinal hypoxia and neovascularizationthat are caused by high metabolic activity in the dark-adapted rodphotoreceptor cell. Merely by way of one example, altering the “dark”visual cycle by administering any one of the compounds described herein,which is an isomerase inhibitor, rhodopsin (i.e., 11-cis retinal bound)may be reduced or depleted, preventing or inhibiting dark adaptation ofrod photoreceptors. This in turn may reduce retinal metabolic demand,attenuating the nighttime risk of retinal ischemia andneovascularization, and thereby inhibiting or slowing retinaldegeneration.

In one embodiment, at least one of the compounds described herein,substructures thereof, and the specific styrenyl compounds recitedherein) that, for example, blocks, reduces, inhibits, or in some mannerattenuates the catalytic activity of a visual cycle isomerase in astatistically or biologically significant manner, may prevent, inhibit,or delay dark adaptation of a rod photoreceptor cell, thereby inhibiting(i.e., reducing, abrogating, preventing, slowing the progression of, ordecreasing in a statistically or biologically significant manner)degeneration of retinal cells (or enhancing survival of retinal cells)of the retina of an eye. In another embodiment, the styrenyl derivativecompounds may prevent or inhibit dark adaptation of a rod photoreceptorcell, thereby reducing ischemia (i.e., decreasing, preventing,inhibiting, slowing the progression of ischemia in a statistically orbiologically significant manner). In yet another embodiment, any one ofthe styrenyl compounds described herein may prevent dark adaptation of arod photoreceptor cell, thereby inhibiting neovascularization in theretina of an eye. Accordingly, methods are provided herein forinhibiting retinal cell degeneration, for inhibiting neovascularizationin the retina of an eye of a subject, and for reducing ischemia in aneye of a subject wherein the methods comprise administering at least onestyrenyl compound described herein, under conditions and at a timesufficient to prevent, inhibit, or delay dark adaptation of a rodphotoreceptor cell. These methods and compositions are therefore usefulfor treating an ophthalmic disease or disorder including, but notlimited to, diabetic retinopathy, diabetic maculopathy, retinal bloodvessel occlusion, retinopathy of prematurity, or ischemia reperfusionrelated retinal injury.

The styrenyl compounds described herein, substructures thereof, and thespecific styrenyl compounds recited herein) may prevent (i.e., delay,slow, inhibit, or decrease) recovery of the visual pigment chromophore,which may prevent or inhibit or retard the formation of retinals and mayincrease the level of retinyl esters, which perturbs the visual cycle,inhibiting regeneration of rhodopsin, and which prevents, slows, delaysor inhibits dark adaptation of a rod photoreceptor cell. In certainembodiments, when dark adaptation of rod photoreceptor cells isprevented in the presence of the compound, dark adaptation issubstantially prevented, and the number or percent of rod photoreceptorcells that are rhodopsin-depleted or light adapted is increased comparedwith the number or percent of cells that are rhodopsin-depleted orlight-adapted in the absence of the agent. Thus, in certain embodimentswhen dark adaptation of rod photoreceptor cells is prevented (i.e.,substantially prevented), only at least 2% of rod photoreceptor cellsare dark-adapted, similar to the percent or number of cells that are ina dark-adapted state during normal, light conditions. In other certainembodiments, at least 5-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, or60-70% of rod photoreceptor cells are dark-adapted after administrationof an agent. In other embodiments, the compound acts to delay darkadaptation, and in the presence of the compound dark adaptation of rodphotoreceptor cells may be delayed 30 minutes, one hour, two hours,three hours, or four hours compared to dark adaptation of rodphotoreceptors in the absence of the compound. By contrast, when astyrenyl compound is administered such that the compound effectivelyinhibits isomerization of substrate during light-adapted conditions, thecompound is administered in such a manner to minimize the percent of rodphotoreceptor cells that are dark-adapted, for example, only 2%, 5%,10%, 20%, or 25% of rod photoreceptors are dark-adapted (see e.g., U.S.Patent Application Publication No. 2006/0069078; Patent Application No.PCT/US2007/002330).

In the retina in the presence of at least one styrenyl compound,regeneration of rhodopsin in a rod photoreceptor cell may be inhibitedor the rate of regeneration may be reduced (i.e., inhibited, reduced, ordecreased in a statistically or biologically significant manner), atleast in part, by preventing the formation of retinals, reducing thelevel of retinals, and/or increasing the level of retinyl esters. Todetermine the level of regeneration of rhodopsin in a rod photoreceptorcell, the level of regeneration of rhodopsin (which may be called afirst level) may be determined prior to permitting contact between thecompound and the retina (i.e., prior to administration of the agent).After a time sufficient for the compound and the retina and cells of theretina to interact, (i.e., after administration of the compound), thelevel of regeneration of rhodopsin (which may be called a second level)may be determined. A decrease in the second level compared with thefirst level indicates that the compound inhibits regeneration ofrhodopsin. The level of rhodopsin generation may be determined aftereach dose, or after any number of doses, and ongoing throughout thetherapeutic regimen to characterize the effect of the agent onregeneration of rhodopsin.

In certain embodiments, the subject in need of the treatments describedherein, may have a disease or disorder that results in or causesimpairment of the capability of rod photoreceptors to regeneraterhodopsin in the retina. By way of example, inhibition of rhodopsinregeneration (or reduction of the rate of rhodopsin regeneration) may besymptomatic in patients with diabetes. In addition to determining thelevel of regeneration of rhodopsin in the subject who has diabetesbefore and after administration of a styrenyl compound described herein,the effect of the compound may also be characterized by comparinginhibition of rhodopsin regeneration in a first subject (or a firstgroup or plurality of subjects) to whom the compound is administered, toa second subject (or second group or plurality of subjects) who hasdiabetes but who does not receive the agent.

In another embodiment, a method is provided for preventing or inhibitingdark adaptation of a rod photoreceptor cell (or a plurality of rodphotoreceptor cells) in a retina comprising contacting the retina and atleast one of the styrenyl compounds described herein, substructuresthereof, and the specific styrenyl compounds recited herein), underconditions and at a time sufficient to permit interaction between theagent and an isomerase present in a retinal cell (such as an RPE cell).A first level of 11-cis-retinal in a rod photoreceptor cell in thepresence of the compound may be determined and compared to a secondlevel of 11-cis-retinal in a rod photoreceptor cell in the absence ofthe compound. Prevention or inhibition of dark adaptation of the rodphotoreceptor cell is indicated when the first level of 11-cis-retinalis less than the second level of 11-cis-retinal.

Inhibiting regeneration of rhodopsin may also include increasing thelevel of 11-cis-retinyl esters present in the RPE cell in the presenceof the compound compared with the level of 11-cis-retinyl esters presentin the RPE cell in the absence of the compound (i.e., prior toadministration of the agent). A two-photon imaging technique may be usedto view and analyze retinosome structures in the RPE, which structuresare believed to store retinyl esters (see, e.g., Imanishi et al., J CellBiol. 164:373-83 (2004), Epub 2004 Jan. 26). A first level of retinylesters may be determined prior to administration of the compound, and asecond level of retinyl esters may be determined after administration ofa first dose or any subsequent dose, wherein an increase in the secondlevel compared to the first level indicates that the compound inhibitsregeneration of rhodopsin.

Retinyl esters may be analyzed by gradient HPLC according to methodspracticed in the art (see, for example, Mata et al., Neuron 36:69-80(2002); Trevino et al. J. Exp. Biol. 208:4151-57 (2005)). To measure11-cis and all-trans retinals, retinoids may be extracted by aformaldehyde method (see, e.g., Suzuki et al., Vis. Res. 28:1061-70(1988); Okajima and Pepperberg, Exp. Eye Res. 65:331-40 (1997)) or by ahydroxylamine method (see, e.g., Groenendijk et al., Biochim. Biophys.Acta. 617:430-38 (1980)) before being analyzed on isocratic HPLC (see,e.g., Trevino et al., supra). The retinoids may be monitoredspectrophotometrically (see, e.g., Maeda et al., J. Neurochem.85:944-956 (2003); Van Hooser et al., J Biol. Chem. 277:19173-82(2002)).

In another embodiment of the methods described herein for treating anophthalmic disease or disorder, for inhibiting retinal cell degeneration(or enhancing retinal cell survival), for inhibiting neovascularization,and for reducing ischemia in the retina, preventing or inhibiting darkadaptation of a rod photoreceptor cell in the retina comprisesincreasing the level of apo-rhodopsin (also called opsin) in thephotoreceptor cell. The total level of the visual pigment approximatesthe sum of rhodopsin and apo-rhodopsin and the total level remainsconstant. Therefore, preventing, delaying, or inhibiting dark adaptationof the rod photoreceptor cell may alter the ratio of apo-rhodopsin torhodopsin. In particular embodiments, preventing, delaying, orinhibiting dark adaptation by administering a styrenyl compounddescribed herein may increase the ratio of the level of apo-rhodopsin tothe level of rhodopsin compared to the ratio in the absence of the agent(for example, prior to administration of the agent). An increase in theratio (i.e., a statistically or biologically significant increase) ofapo-rhodopsin to rhodopsin indicates that the percent or number of rodphotoreceptor cells that are rhodopsin-depleted is increased and thatthe percent or number of rod photoreceptor cells that are dark-adaptedis decreased. The ratio of apo-rhodopsin to rhodopsin may be determinedthroughout the course of therapy to monitor the effect of the agent.

Determining or characterizing the capability of compound to prevent,delay, or inhibit dark adaptation of a rod photoreceptor cell may bedetermined in animal model studies. The level of rhodopsin and the ratioof apo-rhodopsin to rhodopsin may be determined prior to administration(which may be called a first level or first ratio, respectively) of theagent and then after administration of a first or any subsequent dose ofthe agent (which may be called a second level or second ratio,respectively) to determine and to demonstrate that the level ofapo-rhodopsin is greater than the level of apo-rhodopsin in the retinaof animals that did not receive the agent. The level of rhodopsin in rodphotoreceptor cells may be performed according to methods practiced inthe art and provided herein (see, e.g., Example 114).

Also provided herein are methods for inhibiting (reducing, slowing,preventing) degeneration and enhancing retinal neuronal cell survival(or prolonging cell viability) comprising administering to a subject acomposition comprising a pharmaceutically acceptable carrier and astyrenyl derivative compound described in detail herein, including acompound having any one of the structures set forth herein,substructures thereof, and specific styrenyl compounds recited herein. Aretinal neuronal cell includes a photoreceptor cell, a bipolar cell, ahorizontal cell, a ganglion cell, and an amacrine cell. In anotherembodiment, methods are provided for enhancing survival or inhibitingdegeneration of a mature retinal cell such as a RPE cell or a Müllerglial cell. In another embodiment, a method for preventing or inhibitingphotoreceptor degeneration in an eye of a subject or a method forrestoring photoreceptor function in an eye of a subject is provided thatcomprises administering to the subject a composition comprising astyrenyl derivative compound as described herein and a pharmaceuticallyor acceptable carrier (i.e., excipient or vehicle). Such methodscomprise administering to a subject a pharmaceutically acceptableexcipient and a styrenyl derivative compound described herein, includinga compound having any one of the structures set forth herein orsubstructures thereof described herein. Without wishing to be bound bytheory, the compounds described herein may inhibit an isomerization stepof the retinoid cycle and/or may slow chromophore flux in a retinoidcycle in the eye.

The ophthalmic disease may result, at least in part, from lipofuscinpigment(s) accumulation and/or from accumulation ofN-retinylidene-N-retinylethanolamine (A2E) in the eye. Accordingly, incertain embodiments, methods are provided for inhibiting or preventingaccumulation of lipofuscin pigment(s) and/or A2E in the eye of asubject. These methods comprise administering to the subject acomposition comprising a pharmaceutically acceptable carrier and astyrenyl compound as described in detail herein, including a compoundhaving the structure as set forth herein or substructures thereof.

A styrenyl compound can be administered to a subject who has an excessof a retinoid in an eye (e.g., an excess of 11-cis-retinol or11-cis-retinal), an excess of retinoid waste products or intermediatesin the recycling of all-trans-retinal, or the like. Methods describedherein and practiced in the art may be used to determine whether thelevel of one or more endogenous retinoids in a subject are altered(increased or decreased in a statistically significant or biologicallysignificant manner) during or after administration of any one of thecompounds described herein. As described in greater detail herein,rhodopsin, which is composed of the protein opsin and retinal (a vitaminA form), is located in the membrane of the photoreceptor cell in theretina of the eye and catalyzes the only light-sensitive step in vision.The 11-cis-retinal chromophore lies in a pocket of the protein and isisomerized to all-trans retinal when light is absorbed. Theisomerization of retinal leads to a change of the shape of rhodopsin,which triggers a cascade of reactions that lead to a nerve impulse thatis transmitted to the brain by the optic nerve.

Methods of determining endogenous retinoid levels in a vertebrate eye,and an excess or deficiency of such retinoids, are disclosed in, forexample, U.S. Patent Application Publication No: 2005/0159662 (thedisclosure of which is incorporated by reference herein in itsentirety). Other methods of determining endogenous retinoid levels in asubject, which is useful for determining whether levels of suchretinoids are above the normal range, and include for example, analysisby high pressure liquid chromatography (HPLC) of retinoids in abiological sample from a subject. For example, retinoid levels can bedetermined in a biological sample that is a blood sample (which includesserum or plasma) from a subject. A biological sample may also includevitreous fluid, aqueous humor, intraocular fluid, subretinal fluid, ortears.

For example, a blood sample can be obtained from a subject, anddifferent retinoid compounds and levels of one or more of the retinoidcompounds in the sample can be separated and analyzed by normal phasehigh pressure liquid chromatography (HPLC) (e.g., with a HP1100 HPLC anda Beckman, Ultrasphere-Si, 4.6 mm×250 mm column using 10% ethylacetate/90% hexane at a flow rate of 1.4 ml/minute). The retinoids canbe detected by, for example, detection at 325 nm using a diode-arraydetector and HP Chemstation A.03.03 software. An excess in retinoids canbe determined, for example, by comparison of the profile of retinoids(i.e., qualitative, e.g., identity of specific compounds, andquantitative, e.g., the level of each specific compound) in the samplewith a sample from a normal subject. Persons skilled in the art who arefamiliar with such assays and techniques and will readily understandthat appropriate controls are included.

As used herein, increased or excessive levels of endogenous retinoid,such as 11-cis-retinol or 11-cis-retinal, refer to levels of endogenousretinoid higher than those found in a healthy eye of a young vertebrateof the same species. Administration of a styrenyl derivative compoundand reduce or eliminate the requirement for endogenous retinoid. Incertain embodiments, the level of endogenous retinoid may be comparedbefore and after any one or more doses of the agent is administered to asubject to determine the effect of the agent on the level of endogenousretinoids in the subject.

In another embodiment, the methods described herein for treating anophthalmic disease or disorder, for inhibiting neovascularization, andfor reducing ischemia in the retina comprise administering at least oneof the compounds described herein, thereby effecting a decrease inmetabolic demand, which includes effecting a reduction in ATPconsumption and in oxygen consumption in rod photoreceptor cells. Asdescribed herein, consumption of ATP and oxygen in a dark-adapted rodphotoreceptor cell is greater than in rod photoreceptor cells that arelight-adapted or rhodopsin-depleted; thus, use of the agents in themethods described herein may reduce the consumption of ATP in the rodphotoreceptor cells that are prevented, inhibited, or delayed from darkadaptation compared with rod photoreceptor cells that are dark-adapted(such as the cells prior to administration or contact with the agent orcells that are never exposed to the agent).

The methods described herein that may prevent or inhibit dark adaptationof a rod photoreceptor cell may therefore reduce hypoxia (i.e., reducein a statistically or biologically significant manner) in the retina.For example, the level of hypoxia (a first level) may be determinedprior to initiation of the treatment regimen, that is, prior to thefirst dosing of the compound (or a composition, as described herein,comprising the compound). The level of hypoxia (for example, a secondlevel) may be determined after the first dosing, and/or after any secondor subsequent dosing to monitor and characterize hypoxia throughout thetreatment regimen. A decrease (reduction) in the second (or anysubsequent) level of hypoxia compared to the level of hypoxia prior toinitial administration indicates that the compound and the treatmentregiment prevent dark adaptation of the rod photoreceptor cells and maybe used for treating ophthalmic diseases and disorders. Consumption ofoxygen, oxygenation of the retina, and/or hypoxia in the retina may bedetermined using methods practiced in the art. For example, oxygenationof the retina may be determined by measuring the fluorescence offlavoproteins in the retina (see, e.g., U.S. Pat. No. 4,569,354).Another exemplary method is retinal oximetry that measures blood oxygensaturation in the large vessels of the retina near the optic disc. Suchmethods may be used to identify and determine the extent of retinalhypoxia before changes in retinal vessel architecture can be detected.

A subject in need of any one or more of the methods of treatmentdescribed herein may be a human or may be a non-human primate or otheranimal (i.e., veterinary use) who has developed symptoms of anophthalmic disease or disorder or who is at risk for developing anophthalmic disease or disorder. Examples of non-human primates and otheranimals include but are not limited to farm animals, pets, and zooanimals (e.g., horses, cows, buffalo, llamas, goats, rabbits, cats,dogs, chimpanzees, orangutans, gorillas, monkeys, elephants, bears,large cats, etc.).

Retinal Cells

The retina of the eye is a thin, delicate layer of nervous tissue. Themajor landmarks of the retina are the area centralis in the posteriorportion of the eye and the peripheral retina in the anterior portion ofthe eye. The retina is thickest near the posterior sections and becomesthinner near the periphery. The area centralis is located in theposterior retina and contains the fovea and foveola and, in primates,contains the macula. The foveola contains the area of maximal conedensity and, thus, imparts the highest visual acuity in the retina. Thefoveola is contained within the fovea, which is contained within themacula.

The peripheral or anterior portion of the retina increases the field ofvision. The peripheral retina extends anterior to the equator of the eyeand is divided into four regions: the near periphery (most posterior),the mid-periphery, the far periphery, and the ora serrata (mostanterior). The ora serrata denotes the termination of the retina.

The term neuron (or nerve cell) as understood in the art and used hereindenotes a cell that arises from neuroepithelial cell precursors. Matureneurons (i.e., fully differentiated cells) display several specificantigenic markers. Neurons may be classified functionally into threegroups: (1) afferent neurons (or sensory neurons) that transmitinformation into the brain for conscious perception and motorcoordination; (2) motor neurons that transmit commands to muscles andglands; and (3) interneurons that are responsible for local circuitry;and (4) projection interneurons that relay information from one regionof the brain to anther region and therefore have long axons.Interneurons process information within specific subregions of the brainand have relatively shorter axons. A neuron typically has four definedregions: the cell body (or soma); an axon; dendrites; and presynapticterminals. The dendrites serve as the primary input of information fromother neural cells. The axon carries the electrical signals that areinitiated in the cell body to other neurons or to effector organs. Atthe presynaptic terminals, the neuron transmits information to anothercell (the postsynaptic cell), which may be another neuron, a musclecell, or a secretory cell.

The retina is composed of several types of neuronal cells. As describedherein, the types of retinal neuronal cells that may be cultured invitro by this method include photoreceptor cells, ganglion cells, andinterneurons such as bipolar cells, horizontal cells, and amacrinecells. Photoreceptors are specialized light-reactive neural cells andcomprise two major classes, rods and cones. Rods are involved inscotopic or dim light vision, whereas photopic or bright light visionoriginates in the cones by the presence of trichromatic pigments. Manyneurodegenerative diseases that result in blindness, such as age-relatedmacular degeneration, genetic macular dystrophies, retinitis pigmentosa,etc, affect photoreceptors.

Extending from their cell bodies, the photoreceptors have twomorphologically distinct regions, the inner and outer segments. Theouter segment lies furthermost from the photoreceptor cell body andcontains disks that convert incoming light energy into electricalimpulses (phototransduction). The outer segment is attached to the innersegment with a very small and fragile cilium. The size and shape of theouter segments vary between rods and cones and are dependent uponposition within the retina. See Hogan, “Retina” in Histology of theHuman Eye: an Atlas and Text Book (Hogan et al. (eds). WB Saunders;Philadelphia, Pa. (1971)); Eye and Orbit, 8^(th) Ed., Bron et al.,(Chapman and Hall, 1997).

Ganglion cells are output neurons that convey information from theretinal interneurons (including horizontal cells, bipolar cells,amacrine cells) to the brain. Bipolar cells are named according to theirmorphology, and receive input from the photoreceptors, connect withamacrine cells, and send output radially to the ganglion cells. Amacrinecells have processes parallel to the plane of the retina and havetypically inhibitory output to ganglion cells. Amacrine cells are oftensubclassified by neurotransmitter or neuromodulator or peptide (such ascalretinin or calbindin) and interact with each other, with bipolarcells, and with photoreceptors. Bipolar cells are retinal interneuronsthat are named according to their morphology; bipolar cells receiveinput from the photoreceptors and sent the input to the ganglion cells.Horizontal cells modulate and transform visual information from largenumbers of photoreceptors and have horizontal integration (whereasbipolar cells relay information radially through the retina).

Other retinal cells that may be present in the retinal cell culturesdescribed herein include glial cells, such as Müller glial cells, andretinal pigment epithelial cells (RPE). Glial cells surround nerve cellbodies and axons. The glial cells do not carry electrical impulses butcontribute to maintenance of normal brain function. Müller glia, thepredominant type of glial cell within the retina, provide structuralsupport of the retina and are involved in the metabolism of the retina(e.g., contribute to regulation of ionic concentrations, degradation ofneurotransmitters, and remove certain metabolites (see, e.g., Kljavin etal., J. Neurosci. 11:2985 (1991))). Müller's fibers (also known assustentacular fibers of retina) are sustentacular neuroglial cells ofthe retina that run through the thickness of the retina from theinternal limiting membrane to the bases of the rods and cones where theyform a row of junctional complexes.

Retinal pigment epithelial (RPE) cells form the outermost layer of theretina, separated from the blood vessel-enriched choroids by Bruch'smembrane. RPE cells are a type of phagocytic epithelial cell, with somefunctions that are macrophage-like, which lies immediately below theretinal photoreceptors. The dorsal surface of the RPE cell is closelyopposed to the ends of the rods, and as discs are shed from the rodouter segment they are internalized and digested by RPE cells. RPE cellsalso produce, store, and transport a variety of factors that contributeto the normal function and survival of photoreceptors. Another functionof RPE cells is to recycle vitamin A as it moves between photoreceptorsand the RPE during light and dark adaptation in the process known as thevisual cycle.

Described herein is an exemplary long-term in vitro cell culture systempermits and promotes the survival in culture of mature retinal cells,including retinal neurons, for at least 2-4 weeks, over 2 months, or foras long as 6 months. The cell culture system may be used for identifyingand characterizing the styrenyl derivative compounds that are useful inthe methods described herein for treating and/or preventing anophthalmic disease or disorder or for preventing or inhibitingaccumulation in the eye of lipofuscin(s) and/or A2E. Retinal cells areisolated from non-embryonic, non-tumorigenic tissue and have not beenimmortalized by any method such as, for example, transformation orinfection with an oncogenic virus. The cell culture system may compriseall the major retinal neuronal cell types (photoreceptors, bipolarcells, horizontal cells, amacrine cells, and ganglion cells), and alsomay include other mature retinal cells such as retinal pigmentepithelial cells and Müller glial cells.

For example, a blood sample can be obtained from a subject, anddifferent retinoid compounds and levels of one or more of the retinoidcompounds in the sample can be separated and analyzed by normal phasehigh pressure liquid chromatography (HPLC) (e.g., with a HP1100 HPLC anda Beckman, Ultrasphere-Si, 4.6 mm×250 mm column using 10% ethylacetate/90% hexane at a flow rate of 1.4 ml/minute). The retinoids canbe detected by, for example, detection at 325 nm using a diode-arraydetector and HP Chemstation A.03.03 software. An excess in retinoids canbe determined, for example, by comparison of the profile of retinoids(i.e., qualitative, e.g., identity of specific compounds, andquantitative, e.g., the level of each specific compound) in the samplewith a sample from a normal subject. Persons skilled in the art who arefamiliar with such assays and techniques and will readily understandthat appropriate controls are included.

As used herein, increased or excessive levels of endogenous retinoid,such as 11-cis-retinol or 11-cis-retinal, refer to levels of endogenousretinoid higher than those found in a healthy eye of a young vertebrateof the same species. Administration of a styrenyl derivative compoundand reduce or eliminate the requirement for endogenous retinoid.

In Vivo and In Vitro Methods for Determining Therapeutic Effectivenessof Compounds

In one embodiment, methods are provided for using the compoundsdescribed herein for enhancing or prolonging retinal cell survival,including retinal neuronal cell survival and RPE cell survival. Alsoprovided herein are methods for inhibiting or preventing degeneration ofa retinal cell, including a retinal neuronal cell (e.g., a photoreceptorcell, an amacrine cell, a horizontal cell, a bipolar cell, and aganglion cell) and other mature retinal cells such as retinal pigmentepithelial cells and Müller glial cells using the compounds describedherein. Such methods comprise, in certain embodiments, administration ofa styrenyl derivative compound as described herein. Such a compound isuseful for enhancing retinal cell survival, including photoreceptor cellsurvival and retinal pigment epithelia survival, inhibiting or slowingdegeneration of a retinal cell, and thus increasing retinal cellviability, which can result in slowing or halting the progression of anophthalmic disease or disorder or retinal injury, which are describedherein.

The effect of a styrenyl derivative compound on retinal cell survival(and/or retinal cell degeneration) may be determined by using cellculture models, animal models, and other methods that are describedherein and practiced by persons skilled in the art. By way of example,and not limitation, such methods and assays include those described inOglivie et al., Exp. Neurol. 161:675-856 (2000); U.S. Pat. No.6,406,840; WO 01/81551; WO 98/12303; U.S. Patent Application No.2002/0009713; WO 00/40699; U.S. Pat. No. 6,117,675; U.S. Pat. No.5,736,516; WO 99/29279; WO 01/83714; WO 01/42784; U.S. Pat. No.6,183,735; U.S. Pat. No. 6,090,624; WO 01/09327; U.S. Pat. No.5,641,750; U.S. Patent Application Publication No. 2004/0147019; andU.S. Patent Application Publication No. 2005/0059148.

Exemplary methods are described herein and practiced by persons skilledin the art for determining the level of enzymatic activity of a visualcycle isomerase in the presence of any one of the compounds describedherein. A compound that decreases isomerase activity may be useful fortreating an ophthalmic disease or disorder. Thus, methods are providedherein for detecting inhibition of isomerase activity comprisingcontacting (i.e., mixing, combining, or in some manner permitting thecompound and isomerase to interact) a biological sample comprising theisomerase and a styrenyl derivative compound described herein and thendetermining the level of enzymatic activity of the isomerase. A personhaving skill in the art will appreciate that as a control, the level ofactivity of the isomerase in the absence of a compound or in thepresence of a compound known not to alter the enzymatic activity of theisomerase can be determined and compared to the level of activity in thepresence of the compound. A decrease in the level of isomerase activityin the presence of the compound compared to the level of isomeraseactivity in the absence of the compound indicates that the compound maybe useful for treating an ophthalmic disease or disorder, such asage-related macular degeneration or Stargardt's disease. A decrease inthe level of isomerase activity in the presence of the compound comparedto the level of isomerase activity in the absence of the compoundindicates that the compound may also be useful in the methods describedherein for inhibiting or preventing dark adaptation, inhibitingneovascularization and reducing hypoxia and thus useful for treating anophthalmic disease or disorder, for example, diabetic retinopathy,diabetic maculopathy, retinal blood vessel occlusion, retinopathy ofprematurity, or ischemia reperfusion related retinal injury.

The capability of a styrenyl compound described herein to inhibit or toprevent dark adaptation of a rod photoreceptor cell by inhibitingregeneration of rhodopsin may be determined by in vitro assays and/or invivo animal models. By way of example, inhibition of regeneration may bedetermined in a mouse model in which a diabetes-like condition isinduced chemically or in a diabetic mouse model (see, e.g., Phipps etal., Invest. Ophthalmol. Vis. Sci. 47:3187-94 (2006); Ramsey et al.,Invest. Ophthalmol. Vis. Sci. 47:5116-24 (2006)). The level of rhodopsin(a first level) may be determined (for example, spectrophotometrically)in the retina of animals prior to administration of the agent andcompared with the level (a second level) of rhodopsin measured in theretina of animals after administration of the agent. A decrease in thesecond level of rhodopsin compared with the first level of rhodopsinindicates that the agent inhibits regeneration of rhodopsin. Theappropriate controls and study design to determine whether regenerationof rhodopsin is inhibited in a statistically significant or biologicallysignificant manner can be readily determined and implemented by personsskilled in the art.

Methods and techniques for determining or characterizing the effect ofany one of the compounds described herein on dark adaptation andrhodopsin regeneration in rod photoreceptor cells in a mammal, includinga human, may be performed according to procedures described herein andpracticed in the art. For example, detection of a visual stimulus afterexposure to light (i.e., photobleaching) versus time in darkness may bedetermined before administration of the first dose of the compound andat a time after the first dose and/or any subsequent dose. A secondmethod for determining prevention or inhibition of dark adaptation bythe rod photoreceptor cells includes measurement of the amplitude of atleast one, at least two, at least three, or more electroretinogramcomponents, which include, for example, the a-wave and the b-wave. See,for example, Lamb et al., supra; Asi et al., Documenta Ophthalmologica79:125-39 (1992).

Inhibiting regeneration of rhodopsin by a styrenyl compound describedherein may comprise reducing the level of the chromophore,11-cis-retinal, that is produced and present in the RPE cell, andconsequently reducing the level of 11-cis-retinal that is present in thephotoreceptor cell. Thus, the compound, when permitted to contact theretina under suitable conditions and at a time sufficient to preventdark adaptation of a rod photoreceptor cell and to inhibit regenerationof rhodopsin in the rod photoreceptor cell, effects a reduction in thelevel of 11-cis-retinal in a rod photoreceptor cell (i.e., astatistically significant or biologically significant reduction). Thatis, the level of 11-cis retinal in a rod photoreceptor cell is greaterprior to administration of the compound when compared with the level of11-cis-retinal in the photoreceptor cell after the first and/or anysubsequent administration of the compound. A first level of11-cis-retinal may be determined prior to administration of thecompound, and a second level of 11-cis-retinal may be determined afteradministration of a first dose or any subsequent dose to monitor theeffect of the compound. A decrease in the second level compared to thefirst level indicates that the compound inhibits regeneration ofrhodopsin and thus inhibits or prevents dark adaptation of the rodphotoreceptor cells.

An exemplary method for determining or characterizing the capability ofa styrenyl compound to reduce retinal hypoxia includes measuring thelevel of retinal oxygenation, for example, by Magnetic Resonance Imaging(MRI) to measure changes in oxygen pressure (see, e.g., Luan et al.,Invest. Ophthalmol. Vis. Sci. 47:320-28 (2006)).

Animal models may be used to characterize and identify compounds thatmay be used to treat retinal diseases and disorders. A recentlydeveloped animal model may be useful for evaluating treatments formacular degeneration has been described by Ambati et al. (Nat. Med.9:1390-97 (2003); Epub 2003 Oct. 19). This animal model is one of only avery few exemplary animal models presently available for evaluating acompound or any molecule for use in treating (including preventing)progression or development of a retinal disease or disorder. Animalmodels in which the ABCR gene, which encodes an ATP-binding cassettetransporter located in the rims of photoreceptor outer segment discs,may be used to evaluate the effect of a compound. Mutations in the ABCRgene are associated with Stargardt's disease, and heterozygous mutationsin ABCR have been associated with AMD. Accordingly, animals have beengenerated with partial or total loss of ABCR function and may used tocharacterize the styrenyl compounds described herein. (See, e.g., Mataet al., Invest. Ophthalmol. Sci. 42:1685-90 (2001); Weng et al., Cell98:13-23 (1999); Mata et al., Proc. Natl. Acad. Sci. USA 97:7154-49(2000); US 2003/0032078; U.S. Pat. No. 6,713,300).

The effect of any one of the compounds described herein may bedetermined in a diabetic retinopathy animal model, such as described inLuan et al. or may be determined in a normal animal model, in which theanimals have been light or dark adapted in the presence and absence ofany one of the compounds described herein. Another exemplary method fordetermining the capability of the agent to reduce retinal hypoxiameasures retinal hypoxia by deposition of a hydroxyprobe (see, e.g., deGooyer et al. (Invest. Ophthalmol. Vis. Sci. 47:5553-60 (2006)). Such atechnique may be performed in an animal model using Rho⁻/Rho⁻ knockoutmice (see de Gooyer et al., supra) in which at least one compounddescribed herein is administered to group(s) of animals in the presenceand absence of the at least one compound, or may be performed in normal,wildtype animals in which at least one compound described herein isadministered to group(s) of animals in the presence and absence of theat least one compound. Other animal models include models fordetermining photoreceptor function, such as rat models that measureelctroretinographic (ERG) oscillatory potentials (see, e.g., Liu et al.,Invest. Ophthalmol. Vis. Sci. 47:5447-52 (2006); Akula et al., Invest.Ophthalmol. Vis. Sci. 48:4351-59 (2007); Liu et al., Invest. Ophthalmol.Vis. Sci. 47:2639-47 (2006); Dembinska et al., Invest. Ophthalmol. Vis.Sci. 43:2481-90 (2002); Penn et al., Invest. Ophthalmol. Vis. Sci.35:3429-35 (1994); Hancock et al., Invest. Ophthalmol. Vis. Sci.45:1002-1008 (2004)).

Accordingly, cell culture methods, such as the method described herein,is particularly useful for determining the effect of a compounddescribed herein on retinal neuronal cell survival. Exemplary cellculture models are described herein and described in detail in U.S.Patent Application Publication No. US 2005-0059148 and U.S. PatentApplication Publication No. US2004-0147019 (which are incorporated byreference in their entirety), which are useful for determining thecapability of a styrenyl derivative compound as described herein toenhance or prolong survival of neuronal cells, particularly retinalneuronal cells, and of retinal pigment epithelial cells, and inhibit,prevent, slow, or retard degeneration of an eye, or the retina orretinal cells thereof, or the RPE, and which compounds are useful fortreating ophthalmic diseases and disorders.

The cell culture model comprises a long-term or extended culture ofmature retinal cells, including retinal neuronal cells (e.g.,photoreceptor cells, amacrine cells, ganglion cells, horizontal cells,and bipolar cells). The cell culture system and methods for producingthe cell culture system provide extended culture of photoreceptor cells.The cell culture system may also comprise retinal pigment epithelial(RPE) cells and Müller glial cells.

The retinal cell culture system may also comprise a cell stressor. Theapplication or the presence of the stressor affects the mature retinalcells, including the retinal neuronal cells, in vitro, in a manner thatis useful for studying disease pathology that is observed in a retinaldisease or disorder. The cell culture model provides an in vitroneuronal cell culture system that will be useful in the identificationand biological testing of a styrenyl derivative compound that issuitable for treatment of neurological diseases or disorders in general,and for treatment of degenerative diseases of the eye and brain inparticular. The ability to maintain primary, in vitro-cultured cellsfrom mature retinal tissue, including retinal neurons over an extendedperiod of time in the presence of a stressor enables examination ofcell-to-cell interactions, selection and analysis of neuroactivecompounds and materials, use of a controlled cell culture system for invitro CNS and ophthalmic tests, and analysis of the effects on singlecells from a consistent retinal cell population.

The cell culture system and the retinal cell stress model comprisecultured mature retinal cells, retinal neurons, and a retinal cellstressor, which may be used for screening and characterizing a styrenylderivative compound that are capable of inducing or stimulating theregeneration of CNS tissue that has been damaged by disease. The cellculture system provides a mature retinal cell culture that is a mixtureof mature retinal neuronal cells and non-neuronal retinal cells. Thecell culture system may comprise all the major retinal neuronal celltypes (photoreceptors, bipolar cells, horizontal cells, amacrine cells,and ganglion cells), and may also include other mature retinal cellssuch as RPE and Müller glial cells. By incorporating these differenttypes of cells into the in vitro culture system, the system essentiallyresembles an “artificial organ” that is more akin to the natural in vivostate of the retina.

Viability of one or more of the mature retinal cell types that areisolated (harvested) from retinal tissue and plated for tissue culturemay be maintained for an extended period of time, for example, from twoweeks up to six months. Viability of the retinal cells may be determinedaccording to methods described herein and known in the art. Retinalneuronal cells, similar to neuronal cells in general, are not activelydividing cells in vivo and thus cell division of retinal neuronal cellswould not necessarily be indicative of viability. An advantage of thecell culture system is the ability to culture amacrine cells,photoreceptors, and associated ganglion projection neurons and othermature retinal cells for extended periods of time, thereby providing anopportunity to determine the effectiveness of a styrenyl derivativecompound described herein for treatment of retinal disease.

The biological source of the retinal cells or retinal tissue may bemammalian (e.g., human, non-human primate, ungulate, rodent, canine,porcine, bovine, or other mammalian source), avian, or from othergenera. Retinal cells including retinal neurons from post-natalnon-human primates, post-natal pigs, or post-natal chickens may be used,but any adult or post-natal retinal tissue may be suitable for use inthis retinal cell culture system.

In certain instances, the cell culture system may provide for robustlong-term survival of retinal cells without inclusion of cells derivedfrom or isolated or purified from non-retinal tissue. Such a cellculture system comprises cells isolated solely from the retina of theeye and thus is substantially free of types of cells from other parts orregions of the eye that are separate from the retina, such as theciliary body, iris, choroid, and vitreous. Other cell culture methodsinclude the addition of non-retinal cells, such as ciliary body celland/or stem cells (which may or may not be retinal stem cells) and/oradditional purified glial cells.

The in vitro retinal cell culture systems described herein may serve asphysiological retinal models that can be used to characterize aspects ofthe physiology of the retina. This physiological retinal model may alsobe used as a broader general neurobiology model. A cell stressor may beincluded in the model cell culture system. A cell stressor, which asdescribed herein is a retinal cell stressor, adversely affects theviability or reduces the viability of one or more of the differentretinal cell types, including types of retinal neuronal cells, in thecell culture system. A person skilled in the art would readilyappreciate and understand that as described herein a retinal cell thatexhibits reduced viability means that the length of time that a retinalcell survives in the cell culture system is reduced or decreased(decreased lifespan) and/or that the retinal cell exhibits a decrease,inhibition, or adverse effect of a biological or biochemical function(e.g., decreased or abnormal metabolism; initiation of apoptosis; etc.)compared with a retinal cell cultured in an appropriate control cellsystem (e.g., the cell culture system described herein in the absence ofthe cell stressor). Reduced viability of a retinal cell may be indicatedby cell death; an alteration or change in cell structure or morphology;induction and/or progression of apoptosis; initiation, enhancement,and/or acceleration of retinal neuronal cell neurodegeneration (orneuronal cell injury).

Methods and techniques for determining cell viability are described indetail herein and are those with which skilled artisans are familiar.These methods and techniques for determining cell viability may be usedfor monitoring the health and status of retinal cells in the cellculture system and for determining the capability of the styrenylderivative compounds described herein to alter (preferably increase,prolong, enhance, improve) retinal cell or retinal pigment epithelialcell viability or retinal cell survival.

The addition of a cell stressor to the cell culture system is useful fordetermining the capability of a styrenyl derivative compound toabrogate, inhibit, eliminate, or lessen the effect of the stressor. Theretinal cell culture system may include a cell stressor that is chemical(e.g., A2E, cigarette smoke concentrate); biological (for example, toxinexposure; beta-amyloid; lipopolysaccharides); or non-chemical, such as aphysical stressor, environmental stressor, or a mechanical force (e.g.,increased pressure or light exposure) (see, e.g., US 2005-0059148).

The retinal cell stressor model system may also include a cell stressorsuch as, but not limited to, a stressor that may be a risk factor in adisease or disorder or that may contribute to the development orprogression of a disease or disorder, including but not limited to,light of varying wavelengths and intensities; A2E; cigarette smokecondensate exposure; oxidative stress (e.g., stress related to thepresence of or exposure to hydrogen peroxide, nitroprusside, Zn++, orFe++); increased pressure (e.g., atmospheric pressure or hydrostaticpressure), glutamate or glutamate agonist (e.g., N-methyl-D-aspartate(NMDA); alpha-amino-3-hydroxy-5-methylisoxazole-4-proprionate (AMPA);kainic acid; quisqualic acid; ibotenic acid; quinolinic acid; aspartate;trans-1-aminocyclopentyl-1,3-dicarboxylate (ACPD)); amino acids (e.g.,aspartate, L-cysteine; beta-N-methylamine-L-alanine); heavy metals (suchas lead); various toxins (for example, mitochondrial toxins (e.g.,malonate, 3-nitroproprionic acid; rotenone, cyanide); MPTP(1-methyl-4-phenyl-1,2,3,6,-tetrahydropyridine), which metabolizes toits active, toxic metabolite MPP+ (1-methyl-4-phenylpryidine));6-hydroxydopamine; alpha-synuclein; protein kinase C activators (e.g.,phorbol myristate acetate); biogenic amino stimulants (for example,methamphetamine, MDMA (3-4 methylenedioxymethamphetamine)); or acombination of one or more stressors. Useful retinal cell stressorsinclude those that mimic a neurodegenerative disease that affects anyone or more of the mature retinal cells described herein. A chronicdisease model is of particular importance because most neurodegenerativediseases are chronic. Through use of this in vitro cell culture system,the earliest events in long-term disease development processes may beidentified because an extended period of time is available for cellularanalysis.

A retinal cell stressor may alter (i.e., increase or decrease in astatistically significant manner) viability of retinal cells such as byaltering survival of retinal cells, including retinal neuronal cells andRPE cells, or by altering neurodegeneration of retinal neuronal cellsand/or RPE cells. Preferably, a retinal cell stressor adversely affectsa retinal neuronal cell or RPE cell such that survival of a retinalneuronal cell or RPE cell is decreased or adversely affected (i.e., thelength of time during which the cells are viable is decreased in thepresence of the stressor) or neurodegeneration (or neuron cell injury)of the cell is increased or enhanced. The stressor may affect only asingle retinal cell type in the retinal cell culture or the stressor mayaffect two, three, four, or more of the different cell types. Forexample, a stressor may alter viability and survival of photoreceptorcells but not affect all the other major cell types (e.g., ganglioncells, amacrine cells, horizontal cells, bipolar cells, RPE, and Müllerglia). Stressors may shorten the survival time of a retinal cell (invivo or in vitro), increase the rapidity or extent of neurodegenerationof a retinal cell, or in some other manner adversely affect theviability, morphology, maturity, or lifespan of the retinal cell.

The effect of a cell stressor (in the presence and absence of a styrenylderivative compound) on the viability of retinal cells in the cellculture system may be determined for one or more of the differentretinal cell types. Determination of cell viability may includeevaluating structure and/or a function of a retinal cell continually atintervals over a length of time or at a particular time point after theretinal cell culture is prepared. Viability or long term survival of oneor more different retinal cell types or one or more different retinalneuronal cell types may be examined according to one or more biochemicalor biological parameters that are indicative of reduced viability, suchas apoptosis or a decrease in a metabolic function, prior to observationof a morphological or structural alteration.

A chemical, biological, or physical cell stressor may reduce viabilityof one or more of the retinal cell types present in the cell culturesystem when the stressor is added to the cell culture under conditionsdescribed herein for maintaining the long-term cell culture.Alternatively, one or more culture conditions may be adjusted so thatthe effect of the stressor on the retinal cells can be more readilyobserved. For example, the concentration or percent of fetal bovineserum may be reduced or eliminated from the cell culture when cells areexposed to a particular cell stressor (see, e.g., US 2005-0059148).Alternatively, retinal cells cultured in media containing serum at aparticular concentration for maintenance of the cells may be abruptlyexposed to media that does not contain any level of serum.

The retinal cell culture may be exposed to a cell stressor for a periodof time that is determined to reduce the viability of one or moreretinal cell types in the retinal cell culture system. The cells may beexposed to a cell stressor immediately upon plating of the retinal cellsafter isolation from retinal tissue. Alternatively, the retinal cellculture may be exposed to a stressor after the culture is established,or any time thereafter. When two or more cell stressors are included inthe retinal cell culture system, each stressor may be added to the cellculture system concurrently and for the same length of time or may beadded separately at different time points for the same length of time orfor differing lengths of time during the culturing of the retinal cellsystem. A styrenyl compound may be added before the retinal cell cultureis exposed to a cell stressor, may be added concurrently with the cellstressor, or may be added after exposure of the retinal cell culture tothe stressor.

Photoreceptors may be identified using antibodies that specifically bindto photoreceptor-specific proteins such as opsins, peripherins, and thelike. Photoreceptors in cell culture may also be identified as amorphologic subset of immunocytochemically labeled cells by using apan-neuronal marker or may be identified morphologically in enhancedcontrast images of live cultures. Outer segments can be detectedmorphologically as attachments to photoreceptors.

Retinal cells including photoreceptors can also be detected byfunctional analysis. For example, electrophysiology methods andtechniques may be used for measuring the response of photoreceptors tolight. Photoreceptors exhibit specific kinetics in a graded response tolight. Calcium-sensitive dyes may also be used to detect gradedresponses to light within cultures containing active photoreceptors. Foranalyzing stress-inducing compounds or potential neurotherapeutics,retinal cell cultures can be processed for immunocytochemistry, andphotoreceptors and/or other retinal cells can be counted manually or bycomputer software using photomicroscopy and imaging techniques. Otherimmunoassays known in the art (e.g., ELISA, immunoblotting, flowcytometry) may also be useful for identifying and characterizing theretinal cells and retinal neuronal cells of the cell culture modelsystem described herein.

The retinal cell culture stress models may also be useful foridentification of both direct and indirect pharmacologic agent effectsby the bioactive agent of interest, such as a styrenyl derivativecompound as described herein. For example, a bioactive agent added tothe cell culture system in the presence of one or more retinal cellstressors may stimulate one cell type in a manner that enhances ordecreases the survival of other cell types. Cell/cell interactions andcell/extracellular component interactions may be important inunderstanding mechanisms of disease and drug function. For example, oneneuronal cell type may secrete trophic factors that affect growth orsurvival of another neuronal cell type (see, e.g., WO 99/29279).

In another embodiment, a styrenyl derivative compound is incorporatedinto screening assays comprising the retinal cell culture stress modelsystem described herein to determine whether and/or to what level ordegree the compound increases viability (i.e., increases in astatistically significant or biologically significant manner) of aplurality of retinal cells. A person skilled in the art would readilyappreciate and understand that as described herein a retinal cell thatexhibits increased viability means that the length of time that aretinal cell survives in the cell culture system is increased (increasedlifespan) and/or that the retinal cell maintains a biological orbiochemical function (normal metabolism and organelle function; lack ofapoptosis; etc.) compared with a retinal cell cultured in an appropriatecontrol cell system (e.g., the cell culture system described herein inthe absence of the compound). Increased viability of a retinal cell maybe indicated by delayed cell death or a reduced number of dead or dyingcells; maintenance of structure and/or morphology; lack of or delayedinitiation of apoptosis; delay, inhibition, slowed progression, and/orabrogation of retinal neuronal cell neurodegeneration or delaying orabrogating or preventing the effects of neuronal cell injury. Methodsand techniques for determining viability of a retinal cell and thuswhether a retinal cell exhibits increased viability are described ingreater detail herein and are known to persons skilled in the art.

In certain embodiments, a method is provided for determining whether astyrenyl derivative compound, enhances survival of photoreceptor cells.One method comprises contacting a retinal cell culture system asdescribed herein with a styrenyl compound under conditions and for atime sufficient to permit interaction between the retinal neuronal cellsand the compound. Enhanced survival (prolonged survival) may be measuredaccording to methods described herein and known in the art, includingdetecting expression of rhodopsin.

The capability of a styrenyl derivative compound to increase retinalcell viability and/or to enhance, promote, or prolong cell survival(that is, to extend the time period in which retinal cells, includingretinal neuronal cells, are viable), and/or impair, inhibit, or impededegeneration as a direct or indirect result of the herein describedstress may be determined by any one of several methods known to thoseskilled in the art. For example, changes in cell morphology in theabsence and presence of the compound may be determined by visualinspection such as by light microscopy, confocal microscopy, or othermicroscopy methods known in the art. Survival of cells can also bedetermined by counting viable and/or nonviable cells, for instanceImmunochemical or immunohistological techniques (such as fixed cellstaining or flow cytometry) may be used to identify and evaluatecytoskeletal structure (e.g., by using antibodies specific forcytoskeletal proteins such as glial fibrillary acidic protein,fibronectin, actin, vimentin, tubulin, or the like) or to evaluateexpression of cell markers as described herein. The effect of a styrenylderivative compound on cell integrity, morphology, and/or survival mayalso be determined by measuring the phosphorylation state of neuronalcell polypeptides, for example, cytoskeletal polypeptides (see, e.g.,Sharma et al., J. Biol. Chem. 274:9600-06 (1999); Li et al., J Neurosci.20:6055-62 (2000)). Cell survival or, alternatively cell death, may alsobe determined according to methods described herein and known in the artfor measuring apoptosis (for example, annexin V binding, DNAfragmentation assays, caspase activation, marker analysis, e.g.,poly(ADP-ribose) polymerase (PARP), etc.).

In the vertebrate eye, for example, a mammalian eye, the formation ofA2E is a light-dependent process and its accumulation leads to a numberof negative effects in the eye. These include destabilization of retinalpigment epithelium (RPE) membranes, sensitization of cells to blue-lightdamage, and impaired degradation of phospholipids. Products of theoxidation of A2E (and A2E related molecules) by molecular oxygen(oxiranes) were shown to induce DNA damage in cultured RPE cells. Allthese factors lead to a gradual decrease in visual acuity and eventuallyto vision loss. If reducing the formation of retinals during visionprocesses were possible, this reduction would lead to decreased amountsof A2E in the eye. Without wishing to be bound by theory, decreasedaccumulation of A2E may reduce or delay degenerative processes in theRPE and retina and thus may slow down or prevent vision loss in dry AMDand Stargardt's Disease.

In another embodiment, methods are provided for treating and/orpreventing degenerative diseases and disorders, includingneurodegenerative retinal diseases and ophthalmic diseases as describedherein. A subject in need of such treatment may be a human or non-humanprimate or other animal who has developed symptoms of a degenerativeretinal disease or who is at risk for developing a degenerative retinaldisease. As described herein a method is provided for treating (whichincludes preventing or prophylaxis) an ophthalmic disease or disorder byadministrating to a subject a composition comprising a pharmaceuticallyacceptable carrier and a styrenyl derivative compound. As describedherein, a method is provided for enhancing survival of neuronal cellssuch as retinal neuronal cells, including photoreceptor cells, and/orinhibiting degeneration of retinal neuronal cells by administering thepharmaceutical compositions described herein comprising a styrenylderivative compound.

Enhanced survival (or prolonged or extended survival) of one or moreretinal cell types in the presence of a styrenyl derivative compoundindicates that the compound may be an effective agent for treatment of adegenerative disease, particularly a retinal disease or disorder, andincluding a neurodegenerative retinal disease or disorder. Cell survivaland enhanced cell survival may be determined according to methodsdescribed herein and known to a skilled artisan including viabilityassays and assays for detecting expression of retinal cell markerproteins. For determining enhanced survival of photoreceptor cells,opsins may be detected, for instance, including the protein rhodopsinthat is expressed by rods.

In another embodiment, the subject is being treated for Stargardt'sdisease or Stargardt's macular degeneration. In Stargardt's disease,which is associated with mutations in the ABCA4 (also called ABCR)transporter, the accumulation of all-trans-retinal has been proposed tobe responsible for the formation of a lipofuscin pigment, A2E, which istoxic towards retinal cells and causes retinal degeneration andconsequently loss of vision.

In yet another embodiment, the subject is being treated for age-relatedmacular degeneration (AMD). In various embodiments, AMD can be wet ordry form. In AMD, vision loss primarily occurs when complications latein the disease either cause new blood vessels to grow under the maculaor the macula atrophies. Without intending to be bound by any particulartheory, the accumulation of all-trans-retinal has been proposed to beresponsible for the formation of a lipofuscin pigment,N-retinylidene-N-retinylethanolamine (A2E) and A2E related molecules,which are toxic towards RPE and retinal cells and cause retinaldegeneration and consequently loss of vision.

A neurodegenerative retinal disease or disorder for which the compoundsand methods described herein may be used for treating, curing,preventing, ameliorating the symptoms of, or slowing, inhibiting, orstopping the progression of, is a disease or disorder that leads to oris characterized by retinal neuronal cell loss, which is the cause ofvisual impairment. Such a disease or disorder includes but is notlimited to age-related macular degeneration (including dry-form andwet-form of macular degeneration) and Stargardt's macular dystrophy.

Age-related macular degeneration as described herein is a disorder thataffects the macula (central region of the retina) and results in thedecline and loss of central vision. Age-related macular degenerationoccurs typically in individuals over the age of 55 years. The etiologyof age-related macular degeneration may include both environmentalinfluences and genetic components (see, e.g., Lyengar et al., Am. J Hum.Genet. 74:20-39 (2004) (Epub 2003 Dec. 19); Kenealy et al., Mol. Vis.10:57-61 (2004); Gorin et al., Mol. Vis. 5:29 (1999)). More rarely,macular degeneration occurs in younger individuals, including childrenand infants, and generally, these disorders results from a geneticmutation. Types of juvenile macular degeneration include Stargardt'sdisease (see, e.g., Glazer et al., Ophthalmol. Clin. North Am.15:93-100, viii (2002); Weng et al., Cell 98:13-23 (1999)); Doyne'shoneycomb retinal dystrophy (see, e.g., Kermani et al., Hum. Genet.104:77-82 (1999)); Sorsby's fundus dystrophy, Malattia Levintinese,fundus flavimaculatus, and autosomal dominant hemorrhagic maculardystrophy (see also Seddon et al., Ophthalmology 108:2060-67 (2001);Yates et al., J. Med. Genet. 37:83-7 (2000); Jaakson et al., Hum. Mutat.22:395-403 (2003)).

Geographic atrophy of the RPE is an advanced form of non-neovasculardry-type age-related macular degeneration, and is associated withatrophy of the choriocapillaris, RPE, and retina.

Stargardt's macular degeneration, a recessive inherited disease, is aninherited blinding disease of children. The primary pathologic defect inStargardt's disease is also an accumulation of toxic lipofuscin pigmentssuch as A2E in cells of the retinal pigment epithelium (RPE). Thisaccumulation appears to be responsible for the photoreceptor death andsevere visual loss found in Stargardt's patients. The compoundsdescribed herein may slow the synthesis of 11-cis-retinaldehyde (11 cRALor retinal) and regeneration of -rhodopsin by inhibiting isomerase inthe visual cycle. Light activation of rhodopsin results in its releaseof all-trans-retinal, which constitutes the first reactant in A2Ebiosynthesis. Treatment with styrenyl derivative compounds may inhibitlipofuscin accumulation and thus delay the onset of visual loss inStargardt's and AMD patients without toxic effects that would precludetreatment with a styrenyl derivative compound. The compounds describedherein may be used for effective treatment of other forms of retinal ormacular degeneration associated with lipofuscin accumulation.

Administration of a styrenyl derivative compound to a subject canprevent formation of the lipofuscin pigment, A2E (and A2E relatedmolecules), that is toxic towards retinal cells and causes retinaldegeneration. In certain embodiments, administration of a styrenylderivative compound can lessen the production of waste products, e.g.,lipofuscin pigment, A2E (and A2E related molecules), ameliorate thedevelopment of AMD (e.g., dry-form) and Stargardt's disease, and reduceor slow vision loss (e.g., choroidal neovascularization and/orchorioretinal atrophy). In previous studies, with 13-cis-retinoic acid(Accutane® or Isotretinoin), a drug commonly used for the treatment ofacne and an inhibitor of 11-cis-retinol dehydrogenase, has beenadministered to patients to prevent A2E accumulation in the RPE.However, a major drawback in this proposed treatment is that13-cis-retinoic acid can easily isomerize to all-trans-retinoic acid.All-trans-retinoic acid is a very potent teratogenic compound thatadversely affects cell proliferation and development. Retinoic acid alsoaccumulates in the liver and may be a contributing factor in liverdiseases.

In yet other embodiments, a styrenyl derivative compound is administeredto a subject such as a human with a mutation in the ABCA4 transporter inthe eye. The styrenyl derivative compound can also be administered to anaging subject. As used herein, an aging human subject is typically atleast 45, or at least 50, or at least 60, or at least 65 years old. InStargardt's disease, which is associated with mutations in the ABCA4transporter, the accumulation of all-trans-retinal has been proposed tobe responsible for the formation of a lipofuscin pigment, A2E (and A2Erelated molecules), that is toxic towards retinal cells and causesretinal degeneration and consequently loss of vision. Without wishing tobe bound by theory, a styrenyl derivative compound described herein maybe a strong inhibitor of the isomerase protein involved in the visualcycle. Treating patients with a styrenyl derivative compound asdescribed herein may prevent or slow the formation of A2E (and A2Erelated molecules) and can have protective properties for normal vision.

In other certain embodiments, one or more of the compounds describedherein may be used for treating other ophthalmic diseases or disorders,for example, glaucoma, retinal detachment, hemorrhagic retinopathy,retinitis pigmentosa, an inflammatory retinal disease, proliferativevitreoretinopathy, retinal dystrophy, hereditary optic neuropathy,Sorsby's fundus dystrophy, uveitis, a retinal injury, opticalneuropathy, and retinal disorders associated with otherneurodegenerative diseases such as Alzheimer's disease, multiplesclerosis, Parkinson's disease or other neurodegenerative diseases thataffect brain cells, a retinal disorder associated with viral infection,or other conditions such as AIDS. A retinal disorder also includes lightdamage to the retina that is related to increased light exposure (i.e.,overexposure to light), for example, accidental strong or intense lightexposure during surgery; strong, intense, or prolonged sunlightexposure, such as at a desert or snow covered terrain; during combat,for example, when observing a flare or explosion or from a laser device,and the like. Retinal diseases can be of degenerative ornon-degenerative nature. Non-limiting examples of degenerative retinaldiseases include age-related macular degeneration, and Stargardt'smacular dystrophy. Examples of non-degenerative retinal diseases includebut are not limited hemorrhagic retinopathy, retinitis pigmentosa, opticneuropathy, inflammatory retinal disease, diabetic retinopathy, diabeticmaculopathy, retinal blood vessel occlusion, retinopathy of prematurity,or ischemia reperfusion related retinal injury, proliferativevitreoretinopathy, retinal dystrophy, hereditary optic neuropathy,Sorsby's fundus dystrophy, uveitis, a retinal injury, a retinal disorderassociated with Alzheimer's disease, a retinal disorder associated withmultiple sclerosis, a retinal disorder associated with Parkinson'sdisease, a retinal disorder associated with viral infection, a retinaldisorder related to light overexposure, and a retinal disorderassociated with AIDS.

In other certain embodiments, one or more of the compounds describedherein may be used for treating, curing, preventing, ameliorating thesymptoms of, or slowing, inhibiting, or stopping the progression of,certain ophthalmic diseases and disorders including but not limited todiabetic retinopathy, diabetic maculopathy, diabetic macular edema,retinal ischemia, ischemia-reperfusion related retinal injury, andretinal blood vessel occlusion (including venous occlusion and arterialocclusion).

Diabetic retinopathy is a leading cause of blindness in humans and is acomplication of diabetes. Non-proliferative retinopathy may be mild,moderate, or severe, and if left untreated, the disease can progress toproliferative retinopathy, a more serious form of diabetic retinopathy.Proliferative retinopathy occurs when new blood vessels proliferate inand around the retina. Consequently, bleeding into the vitreous,swelling of the retina, and/or retinal detachment may occur, leading toblindness.

Other ophthalmic diseases and disorders that may be treated using thecompounds, compositions, and methods described herein include diseases,disorders, and conditions that are associated with, exacerbated by, orcaused by ischemia in the retina. Retinal ischemia includes ischemia ofthe inner retina and the outer retina. Retinal ischemia can occur fromeither choroidal or retinal vascular diseases, such as central or branchretinal vision occlusion, sickle cell retinopathy, collagen vasculardiseases and thrombocytopenic purpura, Eales disease, and systemic lupuserythematosus.

Retinal ischemia may be associated with retinal blood vessel occlusion.In the United States, both branch and central retinal vein occlusionsare the second most common retinal vascular diseases after diabeticretinopathy. About 7% to 10% of patients who have retinal venousocclusive disease in one eye eventually have bilateral disease. Visualfield loss commonly occurs from macular edema, ischemia, or vitreoushemorrhage secondary to disc or retinal neovascularization induced bythe release of vascular endothelial growth factor.

Arteriolosclerosis at sites of retinal arteriovenous crossings (areas inwhich arteries and veins share a common adventitial sheath) causesconstriction of the wall of a retinal vein by a crossing artery. Theconstriction results in thrombus formation and subsequent occlusion ofthe vein. The blocked vein may lead to macular edema and hemorrhagesecondary to breakdown in the blood-retina barrier in the area drainedby the vein, disruption of circulation with turbulence in venous flow,endothelial damage, and ischemia. Clinically, areas of ischemic retinaappear as feathery white patches called cotton-wool spots.

Branch retinal vein occlusions with abundant ischemia cause acutecentral and paracentral visual field loss corresponding to the locationof the involved retinal quadrants. Retinal neovascularization due toischemia may lead to vitreous hemorrhage and subacute or acute visionloss.

Two types of central retinal vein occlusion, ischemic and nonischemic,may occur depending on whether widespread retinal ischemia is present.Even in the nonischemic type, the macula may still be ischemic.Approximately 25% central retinal vein occlusion is ischemic. Diagnosisof central retinal vein occlusion can usually be made on the basis ofcharacteristic ophthalmoscopic findings, including retinal hemorrhage inall quadrants, dilated and tortuous veins, and cotton-wool spots.Macular edema and foveal ischemia can lead to vision loss. Extracellularfluid increases interstitial pressure, which may result in areas ofretinal capillary closure (i.e., patchy ischemic retinal whitening) orocclusion of a cilioretinal artery.

Patients with ischemic central retinal vein occlusion are more likely topresent with a sudden onset of vision loss and have visual acuity ofless than 20/200, a relative afferent pupillary defect, abundantintraretinal hemorrhages, and extensive nonperfusion on fluoresceinangiography. The natural history of ischemic central retinal veinocclusion is associated with poor outcomes: eventually, approximatelytwo-thirds of patients who have ischemic central retinal vein occlusionwill have ocular neovascularization and one-third will have neovascularglaucoma. The latter condition is a severe type of glaucoma that maylead to rapid visual field and vision loss, epithelial edema of thecornea with secondary epithelial erosion and predisposition to bacterialkeratitis, severe pain, nausea and vomiting, and, eventually, phthisisbulbi (atrophy of the globe with no light perception).

As used herein, a patient (or subject) may be any mammal, including ahuman, that may have or be afflicted with a neurodegenerative disease orcondition, including an ophthalmic disease or disorder, or that may befree of detectable disease. Accordingly, the treatment may beadministered to a subject who has an existing disease, or the treatmentmay be prophylactic, administered to a subject who is at risk fordeveloping the disease or condition. Treating or treatment refers to anyindicia of success in the treatment or amelioration of an injury,pathology or condition, including any objective or subjective parametersuch as abatement; remission; diminishing of symptoms or making theinjury, pathology, or condition more tolerable to the patient; slowingin the rate of degeneration or decline; making the final point ofdegeneration less debilitating; or improving a subject's physical ormental well-being.

The treatment or amelioration of symptoms can be based on objective orsubjective parameters; including the results of a physical examination.The term “therapeutic effect” refers to the reduction, elimination, orprevention of the disease, symptoms of the disease, or sequelae of thedisease in the subject. Treatment includes restoring or improvingretinal neuronal cell functions (including photoreceptor function) in avertebrate visual system, for example, such as visual acuity and visualfield testing etc., as measured over time (e.g., as measured in weeks ormonths). Treatment also includes stabilizing disease progression (i.e.,slowing, minimizing, or halting the progression of an ophthalmic diseaseand associated symptoms) and minimizing additional degeneration of avertebrate visual system. Treatment also includes prophylaxis and refersto the administration of a styrenyl derivative compound to a subject toprevent degeneration or further degeneration or deterioration or furtherdeterioration of the vertebrate visual system of the subject and toprevent or inhibit development of the disease and/or related symptomsand sequelae. The term treating also includes the administration of thecompounds or agents described herein to treat pain, hyperalgesia,allodynia, or nociceptive events and to prevent or delay, to alleviate,or to arrest or inhibit development of the symptoms or conditionsassociated with pain, hyperalgesia, allodynia, nociceptive events, orother disorders.

Various methods and techniques practiced by a person skilled in themedical and ophthalmological arts to determine and evaluate a diseasestate and/or to monitor and assess a therapeutic regimen include, forexample, fluorescein angiogram, fundus photography, indocyanine greendye tracking of the choroidal circulatory system, opthalmoscopy, opticalcoherence tomography (OCT), electroretinography, and visual acuitytesting.

A fluorescein angiogram involves injecting a fluorescein dyeintravenously and then observing any leakage of the dye as it circulatesthrough the eye. Intravenous injection of indocyanine green dye may alsobe used to determine if vessels in the eye are compromised, particularlyin the choroidal circulatory system that is just behind the retina.Fundus photography may be used for examining the optic nerve, macula,blood vessels, retina, and the vitreous. Microaneurysms are visiblelesions in diabetic retinopathy that may be detected in digital fundusimages early in the disease (see, e.g., U.S. Patent ApplicationPublication No. 2007/0002275). An ophthalmoscope may be used to examinethe retina and vitreous. Opthalmoscopy is usually performed with dilatedpupils, to allow the best view inside the eye. Two types ofophthalmoscopes may be used: direct and indirect. The directophthalmoscope is generally used to view the optic nerve and the centralretina. The periphery, or entire retina, may be viewed by using anindirect ophthalmoscope. Optical coherence tomography (OCT) produceshigh resolution, high speed, non-invasive, cross-sectional images ofbody tissue. OCT is noninvasive and provides detection of microscopicearly signs of disruption in tissues.

A subject or patient refers to any vertebrate or mammalian patient orsubject to whom the compositions described herein can be administered.The term “vertebrate” or “mammal” includes humans and non-humanprimates, as well as experimental animals such as rabbits, rats, andmice, and other animals, such as domestic pets (such as cats, dogs,horses), farm animals, and zoo animals. Subjects in need of treatmentusing the methods described herein may be identified according toaccepted screening methods in the medical art that are employed todetermine risk factors or symptoms associated with an ophthalmic diseaseor condition described herein or to determine the status of an existingophthalmic disease or condition in a subject. These and other routinemethods allow the clinician to select patients in need of therapy usingthe methods and formulations described herein.

Pharmaceutical Compositions

In certain embodiments, a styrenyl derivative compound described hereinmay be administered as a pure chemical. In other embodiments, thestyrenyl derivative compound can be combined with a pharmaceuticallysuitable or acceptable carrier (also referred to herein as apharmaceutically suitable (or acceptable) excipient, physiologicallysuitable (or acceptable) excipient, or physiologically suitable (oracceptable) carrier) selected on the basis of a chosen route ofadministration and standard pharmaceutical practice as described, forexample, in Remington: The Science and Practice of Pharmacy (Gennaro,21^(st) Ed. Mack Pub. Co., Easton, Pa. (2005)), the disclosure of whichis hereby incorporated herein by reference, in its entirety.

Accordingly, provided herein is a pharmaceutical composition comprisingone or more styrenyl derivative compounds, or a stereoisomer, prodrug,pharmaceutically acceptable salt, hydrate, solvate, acid salt hydrate,N-oxide or isomorphic crystalline form thereof, of a compound describedherein, together with one or more pharmaceutically acceptable carrierstherefore and, optionally, other therapeutic and/or prophylacticingredients. The carrier(s) is acceptable or suitable if the carrier iscompatible with the other ingredients of the composition and notdeleterious to the recipient of the composition. A pharmaceuticallyacceptable or suitable composition includes an ophthalmologicallysuitable or acceptable composition.

Thus, another embodiment provides a pharmaceutical compositioncomprising a pharmaceutically acceptable excipient and a compound havinga structure of Formula (I):

as an isolated E or Z stereoisomer or a mixture of E and Zstereoisomers, as a tautomer or a mixture of tautomers, or as apharmaceutically acceptable salt, hydrate, solvate, N-oxide or prodrugthereof, wherein:

prodrug thereof, wherein:

R₁ and R₂ are each the same or different and independently hydrogen oralkyl;

R₃, R₄, R₅ and R₆ are each the same or different and independentlyhydrogen, halogen, —OR₁₂, alkyl or fluoroalkyl;

R₇ and R₈ are each the same or different and independently hydrogen oralkyl;

R₉ is hydrogen, alkyl, carbocyclyl or —C(═O)R₁₃;

R₁₀ is hydrogen or alkyl; or

R₉ and R₁₀, together with the nitrogen atom to which they are attached,form an N-heterocyclyl;

R₁₁ is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;

R₁₂ is hydrogen or alkyl;

R₁₃ is alkyl, carbocyclyl or aryl;

W is —C(R₁₄)(R₁₅)—, —O—, —S—, —S(═O)—, —S(═O)₂— or —N(R₁₂)—;

Y is —C(R₁₆)(R₁₇)—;

R₁₄ and R₁₅ are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR₁₂, —NR₁₈R₁₉ or carbocyclyl; or

R₁₄ and R₁₅ form an oxo;

R₁₆ and R₁₇ are each the same or different and independently hydrogen,halogen, alkyl, fluoroalkyl, —OR₁₂, —NR₁₈R₁₉ or carbocyclyl; or

R₁₆ and R₁₇ form an oxo; or

R₁₄ and R₁₆ together form a direct bond to provide a double bondconnecting W and Y; or

R₁₄ and R₁₆ together form a direct bond, and R₁₅ and R₁₇ together form adirect bond to provide a triple bond connecting W and Y;

R₁₈ and R₁₉ are each the same or different and independently hydrogen,alkyl, carbocyclyl, or —C(═O)R₁₃,

In certain embodiments, the pharmaceutical composition comprises acompound of Formula (I), as defined herein, provided that when R₁₁ isphenyl, the compound is not:

-   2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]acetamide;-   (2S,3R)-amino-3-hydroxy-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)-ethenyl]phenyl]-butanamide;-   L-glutamic acid,    1-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]ester;-   glycine, 3-hydroxy-5-[(1E)-2(4-hydroxyphenyl)ethenyl]phenyl ester;-   (2S)-2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]propanamide;-   (2S)-2-amino-3-hydroxy-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]propanamide;-   (2S)-2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]-4-methyl-pentanamide;-   (2S)-2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]-3-methyl-butanamide;    or-   2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenylbutanamide.

Various embodiments further provide pharmaceutical compositionscomprising a pharmaceutically acceptable excipient and a compound of anyone of Formulae (I), (II), (IIa), (IIb), (III) and (IIIa):

wherein, t, p, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, R₉, R₁₀, R₁₄, R₁₅, R₁₆,R₁₇, R₂₀, R₂₁, W and Y are as defined above and herein. Specificembodiments provided herein include pharmaceutical compositionscomprising a pharmaceutically acceptable excipient and a compounddescribed in any one of Tables 1-11 provided herein.

A pharmaceutical composition (e.g., for oral administration or deliveryby injection, or combined devices, or for application as an eye drop)may be in the form of a liquid, semi-solid, or solid. A liquidpharmaceutical composition may include, for example, one or more of thefollowing: sterile diluents such as water for injection, salinesolution, preferably physiological saline, Ringer's solution, isotonicsodium chloride, fixed oils that may serve as the solvent or suspendingmedium, polyethylene glycols, glycerin, propylene glycol or othersolvents; antibacterial agents; antioxidants; chelating agents; buffersand agents for the adjustment of tonicity such as sodium chloride ordextrose. A parenteral preparation can be enclosed in ampoules,disposable syringes or multiple dose vials made of glass or plastic. Theuse of physiological saline is preferred, and an injectablepharmaceutical composition or a composition that is delivered ocularlyis preferably sterile.

A styrenyl derivative compound can be administered to human or othernonhuman vertebrates. In certain embodiments, the compound issubstantially pure, in that it contains less than about 5% or less thanabout 1%, or less than about 0.1%, of other organic small molecules,such as contaminating intermediates or by-products that are created, forexample, in one or more of the steps of a synthesis method. In otherembodiments, a combination of one or more styrenyl derivative compoundscan be administered.

A styrenyl derivative compound can be delivered to a subject by anysuitable means, including, for example, orally, parenterally,intraocularly, intravenously, intraperitoneally, intranasally (or otherdelivery methods to the mucous membranes, for example, of the nose,throat, and bronchial tubes), or by local administration to the eye, orby an intraocular or periocular device. Modes of local administrationcan include, for example, eye drops, intraocular injection, orperiocular injection. Periocular injection typically involves injectionof the synthetic isomerization inhibitor, i.e., styrenyl derivativecompound under the conjunctiva or into the Tennon's space (beneath thefibrous tissue overlying the eye). Intraocular injection typicallyinvolves injection of the styrenyl derivative compound into the vitreous(also called intravitreal injection). In certain embodiments, theadministration is non-invasive, such as by eye drops or oral dosageform, or as a combined device.

A styrenyl derivative compound can be formulated for administrationusing pharmaceutically acceptable (suitable) carriers or vehicles aswell as techniques routinely used in the art. A pharmaceuticallyacceptable or suitable carrier includes an ophthalmologically suitableor acceptable carrier. A carrier is selected according to the solubilityof the styrenyl derivative compound. Suitable ophthalmologicalcompositions include those that are administrable locally to the eye,such as by eye drops, injection or the like. In the case of eye drops,the formulation can also optionally include, for example,ophthalmologically compatible agents such as isotonizing agents such assodium chloride, concentrated glycerin, and the like; buffering agentssuch as sodium phosphate, sodium acetate, and the like; surfactants suchas polyoxyethylene sorbitan mono-oleate (also referred to as Polysorbate80), polyoxyl stearate 40, polyoxyethylene hydrogenated castor oil, andthe like; stabilization agents such as sodium citrate, sodium edentate,and the like; preservatives such as benzalkonium chloride, parabens, andthe like; and other ingredients. Preservatives can be employed, forexample, at a level of from about 0.001 to about 1.0% weight/volume. ThepH of the formulation is usually within the range acceptable toophthalmologic formulations, such as within the range of about pH 4 to8.

For injection, the styrenyl derivative compound can be provided in aninjection grade saline solution, in the form of an injectable liposomesolution, slow-release polymer system, or the like. Intraocular andperiocular injections are known to those skilled in the art and aredescribed in numerous publications including, for example, Spaeth, Ed.,Ophthalmic Surgery: Principles of Practice, W. B. Sanders Co.,Philadelphia, Pa., 85-87, 1990.

For delivery of a composition comprising at least one of the compoundsdescribed herein via a mucosal route, which includes delivery to thenasal passages, throat, and airways, the composition may be delivered inthe form of an aerosol. The compound may be in a liquid or powder formfor intramucosal delivery. For example, the composition may deliveredvia a pressurized aerosol container with a suitable propellant, such asa hydrocarbon propellant (e.g., propane, butane, isobutene). Thecomposition may be delivered via a non-pressurized delivery system suchas a nebulizer or atomizer.

Suitable oral dosage forms include, for example, tablets, pills,sachets, or capsules of hard or soft gelatin, methylcellulose or ofanother suitable material easily dissolved in the digestive tract.Suitable nontoxic solid carriers can be used which include, for example,pharmaceutical grades of mannitol, lactose, starch, magnesium stearate,sodium saccharin, talcum, cellulose, glucose, sucrose, magnesiumcarbonate, and the like. (See, e.g., Remington: The Science and Practiceof Pharmacy (Gennaro, 21^(st) Ed. Mack Pub. Co., Easton, Pa. (2005)).

The styrenyl derivative compounds described herein may be formulated forsustained or slow-release. Such compositions may generally be preparedusing well known technology and administered by, for example, oral,periocular, intraocular, rectal or subcutaneous implantation, or byimplantation at the desired target site. Sustained-release formulationsmay contain an agent dispersed in a carrier matrix and/or containedwithin a reservoir surrounded by a rate controlling membrane. Excipientsfor use within such formulations are biocompatible, and may also bebiodegradable; preferably the formulation provides a relatively constantlevel of active component release. The amount of active compoundcontained within a sustained-release formulation depends upon the siteof implantation, the rate and expected duration of release, and thenature of the condition to be treated or prevented.

Systemic drug absorption of a drug or composition administered via anocular route is known to those skilled in the art (see, e.g., Lee etal., Int. J. Pharm. 233:1-18 (2002)). In one embodiment, a styrenylderivative compound is delivered by a topical ocular delivery method(see, e.g., Curr. Drug Metab. 4:213-22 (2003)). The composition may bein the form of an eye drop, salve, or ointment or the like, such as,aqueous eye drops, aqueous ophthalmic suspensions, non-aqueous eyedrops, and non-aqueous ophthalmic suspensions, gels, ophthalmicointments, etc. For preparing a gel, for example, carboxyvinyl polymer,methyl cellulose, sodium alginate, hydroxypropyl cellulose, ethylenemaleic anhydride polymer and the like can be used.

The dose of the composition comprising at least one of the styrenylderivative compounds described herein may differ, depending upon thepatient's (e.g., human) condition, that is, stage of the disease,general health status, age, and other factors that a person skilled inthe medical art will use to determine dose. When the composition is usedas eye drops, for example, one to several drops per unit dose,preferably 1 or 2 drops (about 50 μl per 1 drop), may be applied about 1to about 6 times daily.

Pharmaceutical compositions may be administered in a manner appropriateto the disease to be treated (or prevented) as determined by personsskilled in the medical arts. An appropriate dose and a suitable durationand frequency of administration will be determined by such factors asthe condition of the patient, the type and severity of the patient'sdisease, the particular form of the active ingredient, and the method ofadministration. In general, an appropriate dose and treatment regimenprovides the composition(s) in an amount sufficient to providetherapeutic and/or prophylactic benefit (e.g., an improved clinicaloutcome, such as more frequent complete or partial remissions, or longerdisease-free and/or overall survival, or a lessening of symptomseverity). For prophylactic use, a dose should be sufficient to prevent,delay the onset of, or diminish the severity of a disease associatedwith degeneration of a retinal cell including neurodegeneration ofretinal neuronal cells, ischemia, and/or neovascularization of theretina. Optimal doses may generally be determined using experimentalmodels and/or clinical trials. The optimal dose may depend upon the bodymass, weight, or blood volume of the patient.

The doses of the styrenyl derivative compounds can be suitably selecteddepending on the clinical status, condition and age of the subject,dosage form and the like. In the case of eye drops, a styrenylderivative compound can be administered, for example, from about 0.01mg, about 0.1 mg, or about 1 mg, to about 25 mg, to about 50 mg, toabout 90 mg per single dose. Eye drops can be administered one or moretimes per day, as needed. In the case of injections, suitable doses canbe, for example, about 0.0001 mg, about 0.001 mg, about 0.01 mg, orabout 0.1 mg to about 10 mg, to about 25 mg, to about 50 mg, or to about90 mg of the styrenyl derivative compound, one to seven times per week.In other embodiments, about 1.0 to about 30 mg of the styrenylderivative compound can be administered one to seven times per week.

Oral doses can typically range from 1.0 to 1000 mg, administered one tofour times, or more, per day. An exemplary dosing range for oraladministration is from 10 to 250 mg one to three times per day. If thecomposition is a liquid formulation, the composition may comprise atleast 0.1% active compound at particular mass or weight (e.g., from 1.0to 1000 mg) per unit volume of carrier, for example, from about 2% toabout 60%.

In certain embodiments, at least one styrenyl compound described hereinmay be administered under conditions and at a time that inhibits orprevents dark adaptation of rod photoreceptor cells. In certainembodiments, the compound is administered to a subject at least 30minutes (half hour), 60 minutes (one hour), 90 minutes (1.5 hour), or120 minutes (2 hours) prior to sleeping. In certain embodiments, thecompound may be administered at night before the subject sleeps. Inother embodiments, a light stimulus may be blocked or removed during theday or under normal light conditions by placing the subject in anenvironment in which light is removed, such as placing the subject in adarkened room or by applying an eye mask over the eyes of the subject.When the light stimulus is removed in such a manner or by other meanscontemplated in the art, the agent may be administered prior tosleeping.

The doses of the compounds that may be administered to prevent orinhibit dark adaptation of a rod photoreceptor cell can be suitablyselected depending on the clinical status, condition and age of thesubject, dosage form and the like. In the case of eye drops, thecompound (or the composition comprising the compound) can beadministered, for example, from about 0.01 mg, about 0.1 mg, or about 1mg, to about 25 mg, to about 50 mg, to about 90 mg per single dose. Inthe case of injections, suitable doses can be, for example, about 0.0001mg, about 0.001 mg, about 0.01 mg, or about 0.1 mg to about 10 mg, toabout 25 mg, to about 50 mg, or to about 90 mg of the compound,administered any number of days between one to seven days per week priorto sleeping or prior to removing the subject from all light sources. Incertain other embodiments, for administration of the compound by eyedrops or injection, the dose is between 1-10 mg (compound)/kg (bodyweight of subject) (i.e., for example, 80-800 mg total per dose for asubject weighing 80 kg). In other embodiments, about 1.0 to about 30 mgof compound can be administered one to seven times per week. Oral dosescan typically range from about 1.0 to about 1000 mg, administered anynumber of days between one to seven days per week. An exemplary dosingrange for oral administration is from about 10 to about 800 mg once perday prior to sleeping. In other embodiments, the composition may bedelivered by intravitreal administration.

Also provided are methods of manufacturing the compounds andpharmaceutical compositions described herein. A composition comprising apharmaceutically acceptable excipient or carrier and at least one of thestyrenyl derivative compounds described herein may be prepared bysynthesizing the compound according to any one of the methods describedherein or practiced in the art and then formulating the compound with apharmaceutically acceptable carrier. Formulation of the composition willbe appropriate and dependent on several factors, including but notlimited to, the delivery route, dose, and stability of the compound.

Other embodiments and uses will be apparent to one skilled in the art inlight of the present disclosures. The following examples are providedmerely as illustrative of various embodiments and shall not be construedto limit the invention in any way.

EXAMPLES

Unless otherwise noted, reagents and solvents were used as received fromcommercial suppliers. Anhydrous solvents and oven-dried glassware wereused for synthetic transformations generally considered sensitive tomoisture and/or oxygen. These transformations were conducted undernitrogen or argon atmosphere. Chromatography was performed on silica gelunless otherwise noted. Flash chromatography conducted with a gradientwas performed on a Biotage chromatography system. Proton and carbonnuclear magnetic resonance spectra were obtained on a Bruker AMX 500spectrometer at 500 or 300 MHz for proton and 125 or 75 MHz for carbon,as noted. Spectra are given in ppm (δ) and coupling constants, J, arereported in Hertz. Tetramethylsilane was used as an internal standardfor proton spectra and the solvent peak was used as the reference peakfor carbon spectra. Mass spectra were obtained on a Finnigan LCQ DuoLCMS ion trap electrospray ionization (ESI) mass spectrometer or aPESCiex API 150EX mass spectrometer using atmospheric chemicalionization (APCI). HPLC analyses were obtained using a Symmetry C18column (250×4.6 mm, Waters) with UV detection at 254 nm using a standardsolvent gradient program (Methods 1, 2 and 4), a Pursuit C18 column(100×4.0 mm, Varian) with UV detection at 254 nm using a standardsolvent gradient program (Method 3), a Hypersil BDS C18 column (250×4.6mm, Phenomenex) with detection at 254 nm using a standard solventgradient program (Methods 5 and 6), a Luna 5μ C18(2) column (250×4.6 mm,Phenomenex) with detection at 254 nm using a standard solvent gradientprogram (Method 7), or a Gemini C18 column (150×4.6 mm, 5μ, Phenomenex)with detection at 220 nm using a standard solvent gradient program(Method 8).

Method 1

Time Flow (min) (mL/min) % A % B 0.0 1.0 90.0 10.0 15.0 1.0 0.0 100.020.0 1.0 0.0 100.0 A = Water with 0.05% Trifluoroacetic Acid B =Acetonitrile with 0.05% Trifluoroacetic Acid

Method 2

Time Flow (min) (mL/min) % A % B 0.0 1.0 70.0 30.0 15.0 1.0 0.0 100.020.0 1.0 0.0 100.0 A = Water with 0.05% Trifluoroacetic Acid B =Acetonitrile with 0.05% Trifluoroacetic Acid

Method 3

Time Flow (min) (mL/min) % A % B 0.0 2.0 75.0 25.0 5.0 2.0 5.0 95.0 9.592.0 5.0 95.0 10.0 2.0 75.0 25.0 A = Water with 0.05% TrifluoroaceticAcid B = Acetonitrile with 0.05% Trifluoroacetic Acid

Method 4

Time Flow (min) (mL/min) % A % B 0.0 1.0 70.0 30.0 15.0 1.0 0.0 100.025.0 1.0 0.0 100.0 A = Water with 0.05% Trifluoroacetic Acid B =Acetonitrile with 0.05% Trifluoroacetic Acid

Method 5

Time Flow (min) (mL/min) % A % B 0.0 1.0 70.0 30.0 15.0 1.0 0.0 100.020.0 1.0 0.0 100.0 A = Water with 0.05% Trifluoroacetic Acid B =Acetonitrile with 0.05% Trifluoroacetic Acid

Method 6

Time Flow (min) (mL/min) % A % B 0.0 1.0 90.0 10.0 15.0 1.0 0.0 100.020.0 1.0 0.0 100.0 A = Water with 0.05% Trifluoroacetic Acid B =Acetonitrile with 0.05% Trifluoroacetic Acid

Method 7

Time Flow (min) (mL/min) % A % B 0.0 1.0 70.0 30.0 17.0 1.0 0.0 100.022.0 1.0 0.0 100.0 25.0 1.0 70.0 30.0 A = Water with 0.05%Trifluoroacetic Acid B = Acetonitrile with 0.05% Trifluoroacetic Acid

Method 8

Time Flow (min) (mL/min) % A % B 0.0 1.0 70.0 30.0 6.0 1.0 20.0 80.0 9.01.0 5.0 95.0 A = Water with 0.05% Trifluoroacetic Acid B = Acetonitrilewith 0.05% Trifluoroacetic Acid

Method 9

Time Flow (min) (mL/min) % A % B 0.0 1.0 90.0 10.0 6.0 1.0 20.0 80.0 9.01.0 5.0 95.0 A = Water with 0.05% Trifluoroacetic Acid B = Acetonitrilewith 0.05% Trifluoroacetic Acid

Method 10

Time Flow (min) (mL/min) % A % B 0.0 1.0 90.0 10.0 10.0 1.0 5.0 95.0 A =Water with 0.05% Trifluoroacetic Acid B = Acetonitrile with 0.05%Trifluoroacetic Acid Column = Gemini C18, 4.6 ×150 mm, 5μGeneral Preparative HPLC Conditions:

Preparative HPLC (Method 1) was performed using a Luna 5μ C18 column(250×21.2 mm, Phenomenex) with UV detection at 254 nm and the solventgradient program:

Preparative HPLC (Method 1)

Time Flow (min) (mL/min) % A % B 0.0 15.0 80.0 20.0 30.0 15.0 0.0 100.0A = Water with 0.05% Trifluoroacetic Acid B = Acetonitrile with 0.05%Trifluoroacetic Acid

Preparative HPLC (Method 2) was performed using a YMC ODA-A column (500mm×30 mm×10μ) at ambient temperature with detection at 220 nm using aninjection volume of 5 mL and a standard solvent gradient program.

Preparative HPLC (Method 2)

Time Flow (min) (mL/min) % A % B 0.0 30 90 10 5.0 30 90 10 25 30 20 8035 30 80 80 A = Water with 0.05% Trifluoroacetic Acid B = Acetonitrilewith 0.05% Trifluoroacetic Acid

Chiral HPLC analyses were obtained using a Chiralpak IA column (4.6mm×250 mm, 5μ) with diode array detection. The eluent was 95%heptane-EtOH containing 0.1% ethanesulfonic acid. The flow rate was 1mL/min and the column temperature was 40° C.

Example 1 Preparation of(E)-3-(3-(2,6-dimethylstyryl)phenyl)propan-1-amine

(E)-3-(3-(2,6-dimethylstyryl)phenyl)propan-1-amine was preparedfollowing the method described in Scheme 1. (See, also, Methods A and J)

Step 1:

To a stirred solution of 2,6-dimethylbenzoic acid (1) (10.0 g, 66.6mmol) in THF (100 mL) at 0° C. was added borane-THF complex (80 mL, 1Msolution in THF, 80.0 mmol) dropwise over 20 min and then the reactionmixture was warmed to room temperature. After 64 h the reaction mixturewas quenched by slow addition of methanol (70 mL) and the resultingsolution concentrated. The residue was suspended in ethyl acetate (300mL) and washed with water (4×50 mL) and brine (50 mL), and the organiclayer was dried (Na₂SO₄), filtered and concentrated. The residue wasdried in vacuo to give 2 (9.10 g, >99%) as a white solid: ¹H NMR (500MHz, CDCl₃) δ 7.13-7.03 (m, 3H), 4.74 (d, J=5.1 Hz, 2H), 2.43 (s, 6H),1.28 (t, J=5.2 Hz, 1H); ESI MS m/z 119 [M+H−H₂O]⁺.

Step 2:

To a stirred solution of triphenylphosphine hydrobromide (22.0 g, 64.0mmol) in MeOH (80 mL) was added a solution of 2 (8.72 g, 64.0 mmol) inmethanol (70 mL) and the reaction mixture stirred at room temperaturefor 48 h. The reaction solution was concentrated under reduced pressure,the residue triturated with a mixture of acetone (20 mL) and diethylether (50 mL). The precipitate was collected by vacuum filtration,washed with diethyl ether (30 mL) and hexanes (30 mL), and dried invacuo to provide 3 (23.0 g, 78%) as a white solid: mp 240-246° C.; ¹HNMR (300 MHz, DMSO-d₆) δ 7.95-7.51 (m, 15H), 7.15 (dt, J=7.7, 2.6 Hz,1H), 6.96 (d, J=7.7 Hz, 2H), 4.94 (d, J=14.6 Hz, 2H), 1.76 (s, 6H); ESIMS m/z 381 [M−Br]⁺; HPLC (Method 5) 97.0% (AUC), t_(R)=13.78 min.

Step 3:

To a stirred suspension of 3 (8.76 g, 19.0 mmol) in THF (60 mL) at −78°C. was added n-butyl lithium (7.8 mL, 2.5M solution in hexanes, 19.5mmol) and the reaction mixture warmed to room temperature. After 30 minthe reaction mixture was again cooled to −78° C., a solution of3-iodobenzaldehyde (4.41 g, 19.0 mmol) in THF (15 mL) was added, andthen the reaction mixture was warmed to room temperature. After 1 hr,the reaction was quenched with saturated aqueous ammonium chloride (50mL) and extracted with ethyl acetate (2×100 mL). The combined organiclayers were concentrated and the resulting residue was dissolved inmethanol (70 mL). To this was then added hexanes (400 mL) and water (30mL) and phases were mixed. The hexanes layer was separated, washed with70% methanol/water (100 mL), dried (Na₂SO₄), filtered and concentrated.The residue was purified by flash column chromatography (silica gel,hexanes) to give 4 (2.12 g, 33%) as a white solid and 5 (1.15 g, 18%) asa colorless oil.

4: ¹H NMR (500 MHz, CDCl₃) δ 7.84 (s, 1H), 7.59 (d, J=7.7 Hz, 1H), 7.44(d, J=7.7 Hz, 1H), 7.10-7.06 (m, 5H), 6.49 (d, J=16.6 Hz, 1H), 2.35 (s,6H).

5: ¹H NMR (500 MHz, CDCl₃) δ 7.44 (d, J=7.8 Hz, 1H), 7.37 (s, 1H), 7.13(t, J=7.5 Hz, 1H), 7.04 (d, J=7.6 Hz, 2H), 6.88 (d, J=7.9 Hz, 1H), 6.81(t, J=7.8 Hz, 1H), 6.59 (d, J=12.2 Hz, 1H), 6.53 (d, J=12.2 Hz, 1H),2.14 (s, 6H).

Step 4

To a stirred solution of 4 (1.86 g, 5.60 mmol) in DMF (5 mL) was addedsodium bicarbonate (1.49 g, 17.7 mmol), tetrabutylammonium chloride(1.58 g, 5.70 mmol), and allyl alcohol (0.683 g, 11.8 mmol) The reactionflask was purged with nitrogen for 10 min and then palladium acetate(0.029 g, 0.130 mmol) was added. The color of the solution changed fromyellow to orange. After purging with nitrogen for an additional 10 minthe solution was stirred under nitrogen at room temperature. After 18 hthe solution was diluted with ethyl acetate (50 mL), the resultingmixture washed with water (25 mL), 5% aqueous lithium chloride solution(25 mL), and brine (25 mL). The organics were dried (MgSO₄), filteredand concentrated to afford a dark oil. Purification by columnchromatography (silica, 0-20% ethyl acetate/hexanes) provided 1.05 g(71%) of 6 as a colorless oil: ¹H NMR (500 MHz, CDCl₃) δ 9.85 (t, J=1.1Hz, 1H), 7.39 (d, J=7.8 Hz, 1H), 7.32-7.29 (m, 2H), 7.12-7.07 (m, 5H),6.57 (d, J=16.6 Hz, 1H), 2.99 (t, J=7.6 Hz, 2H), 2.84-2.38 (m, 2H), 2.37(s, 6H).

Step 5:

To a stirred solution of 6 (0.848 g, 3.20 mmol) in methanol (50 mL) wasadded 7N ammonia in methanol (9.0 mL) and a small scoop of poweredsieves. The flask was stoppered and stirred for 1.5 h, at which timesodium borohydride (0.184 g, 4.90 mmol) was added. The solution wasstirred for an additional 3 h, filtered over diatomaceous earth, thefilter cake rinsed with methanol (25 mL) and the filtrate concentrated.The resulting residue was dissolved in ethyl acetate (50 mL) and thissolution was washed with water (25 mL). The aqueous phase was extractedwith ethyl acetate (2×25 mL), the combined organics washed with brine(25 mL), dried (MgSO₄), filtered and concentrated. Purification bycolumn chromatography (silica, 99:1 to 95:5 methylene chloride/7Nammonia in methanol) afforded 0.368 g (43%) of(E)-3-(3-(2,6-dimethylstyryl)phenyl)propan-1-amine.

The salt was formed as follows:

To a stirred solution of(E)-3-(3-(2,6-dimethylstyryl)phenyl)propan-1-amine (0.296 g, 1.10 mmol)in diethyl ether (15 mL) was added 1N HCl in diethyl ether (1.2 mL, 1.20mmol), resulting in immediate precipitation of a white solid. Thesuspension was stirred for 1.5 h, filtered and dried under vacuum toafford 0.314 g (95%) of(E)-3-(3-(2,6-dimethylstyryl)phenyl)propan-1-amine hydrochloride as alight yellow solid: mp 135-136° C.; ¹H NMR (500 MHz, CD₃OD) δ 7.40 (d,J=11.0 Hz, 2H), 7.31 (t, J=7.6 Hz, 1H), 7.21 (s, 1H), 7.17-7.14 (m, 1H),7.04 (s, 3H), 6.58 (d, J=16.6 Hz, 1H), 2.95 (t, J=7.7 Hz, 1H), 2.75 (t,J=7.6 Hz, 2H), 2.34 (s, 6H), 2.00 (quint, J=7.7 Hz, 2H), 1.90 (m, 2H);¹³C NMR (75 MHz, CD₃OD) δ 142.2, 139.4, 138.2, 137.1, 135.2, 130.0,128.9, 128.7, 128.3, 127.9, 127.4, 125.4, 40.4, 33.5, 30.4, 21.2; ESI MSm/z 266 [M+H]⁺ HPLC (Method 2)>99% (AUC), t_(R)=16.4 min. Calcd forC₁₉H₂₃N.HCl: C, 75.60; H, 8.01; N, 4.64. Found: C, 75.25; H, 7.99; N,4.66.

Example 2 Preparation of(Z)-3-(3-(2,6-dimethylstyryl)phenyl)propan-1-amine

(Z)-3-(3-(2,6-dimethylstyryl)phenyl)propan-1-amine was preparedfollowing the method used in Example 1.

Step 1:

(Z)-3-(3-(2,6-Dimethylstyryl)phenyl)propanal was prepared following themethod described in Example 1. Purification by column chromatography(silica, 0-20% ethyl acetate/hexanes) gave (0.054 g, 26%) of a yellowoil: R_(f) 0.56 (silica gel, 90:10 methylene chloride/7N ammonia inmethanol); ¹H NMR (500 MHz, CD₃OD) δ ¹H NMR (500 MHz, CDCl₃) δ 9.67 (t,J=1.3 Hz, 1H), 7.10 (q, J=7.7 Hz, 2H), 7.06-7.03 (m, 2H), 6.95 (d, J=7.6Hz, 1H), 6.85 (d, J=7.8 Hz, 1H), 6.76 (s, 1H), 6.62 (d, J=12.2 Hz, 1H),6.54 (d, J=12.2 Hz, 1H), 2.75 (t, J=7.6 Hz, 2H), 2.53-2.50 (m, 2H), 2.14(s, 6H);

ESI MS m/z 247 [M+H−H₂O]⁺.

Step 2:

Reductive amination of (Z)-3-(3-(2,6-Dimethylstyryl)phenyl)propanal withammonia following the method described in Example 1 followed bypurification by column chromatography (silica, 99:1 to 95:5 methylenechloride/7N ammonia in methanol), followed by further purification byPreparative HPLC (Method 1) afforded Example 2 as a yellow oil. Yield(0.054 g, 26%): R_(f) 0.56 (silica gel, 90:10 methylene chloride/7Nammonia in methanol); ¹H NMR (500 MHz, CD₃OD) δ 7.10-7.07 (m, 1H),7.04-7.0 (m, 3H), 6.95 (d, J=7.6 Hz, 1H), 6.81 (d, J=7.2 Hz, 1H), 6.78(s, 1H), 6.66 (d, J=12.2 Hz, 1H), 6.53 (d, J=12.2 Hz, 1H), 2.48-2.40 (m,4H), 2.12 (s, 6H); 1.55 (quint, J=7.3 Hz, 2H); ¹³C NMR (75 MHz, CD₃OD) δ143.0, 138.9, 138.5, 136.6, 132.3, 129.7, 129.2, 129.0, 128.5, 128.4,128.0, 126.9, 41.8, 35.2, 33.9, 20.4; ESI MS m/z 266 [M+H]⁺ HPLC (Method2)>99% (AUC), t_(R)=8.19 min. HRMS calcd for C₁₉H₂₃N [M+H]: 266.1908.Found: 266.1909.

Example 3 Preparation of(E)-4-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)morpholine

(E)-4-(3-(3-(2,6-Dimethylstyryl)phenyl)propyl)morpholine was preparedfollowing the method described in Scheme 2.

Step 1:

To a stirred solution of 3-bromobenzaldehyde (7) (3.70 g, 20.0 mmol),acrolein diethyl acetal (7.81 g, 60.0 mmol) and tributylamine (7.41 g,40.0 mmol) in DMF (40 mL) was added tetrabutylammonium chloride (5.56 g,20.0 mmol) followed by palladium(II) acetate (0.135 g, 0.601 mmol) andthe reaction mixture was heated at 90° C. After 27 h the reactionmixture was cooled to room temperature, diluted with 2N hydrochloricacid (20 mL), and extracted with a 1:1 mixture of diethyl ether andhexanes (3×150 mL), the combined extracts were washed with water (3×100mL), dried (Na₂SO₄), filtered and concentrated. The resulting residuewas purified by flash column chromatography (silica gel, 90:10hexanes/ethyl acetate) to give 8 (2.97 g, 72%) as a light yellow oil: ¹HNMR (500 MHz, CDCl₃) δ 10.00 (s, 1H), 7.74-7.45 (m, 4H), 4.13 (q, J=7.1Hz, 2H), 3.04 (t, J=7.6 Hz, 2H), 2.66 (t, J=7.6 Hz, 2H), 1.23 (t, J=7.1Hz, 3H).

Step 2:

Compound 9 was prepared following the procedure described for thesynthesis of Compound 4, as a colorless oil, and as a mixture of cis andtrans isomers. The isomers were not separated at this time. Yield (4.10g, 92%), trans-/cis-isomer ratio 2:1.

trans-isomer: ¹H NMR (500 MHz, CDCl₃) δ 7.36-6.80 (m, 8H), 6.59 (d,J=16.6 Hz, 1H), 4.14 (m, 2H), 2.98 (t, J=7.7 Hz, 2H), 2.66 (t, J=7.7 Hz,2H), 2.37 (s, 6H), 1.23 (m, 3H);

cis-isomer: ¹H NMR (500 MHz, CDCl₃) δ 7.36-6.80 (m, 7H), 6.62 (d, J=12.2Hz, 1H), 6.53 (d, J=12.2 Hz, 1H), 4.09 (m, 2H), 2.75 (t, J=7.7 Hz, 2H),2.40 (t, J=7.7 Hz, 2H), 2.14 (s, 6H), 1.23 (m, 3H); ESI MS m/z 263[M+H−EtOH]⁺.

Step 3:

To a stirred solution of 9 (4.10 g, 13.3 mmol) in methanol (20 mL), THF(20 mL) and water (10 mL) was added lithium hydroxide (0.956 g, 39.9mmol) at room temperature. After 3 h the reaction mixture wasconcentrated, the residue was diluted with brine (20 mL) and theresulting mixture was acidified with 2N hydrochloric acid to pH 2. Theaqueous mixture was extracted with methylene chloride (3×100 mL) and thecombined extracts were dried (Na₂SO₄), filtered and concentrated. Theresulting residue was dried in vacuo to give 10 as a colorless oil.Yield (3.73 g, quant.), trans-/cis-isomer ratio 2:1.

trans-isomer: ¹H NMR (500 MHz, CDCl₃) δ 7.39-6.80 (m, 8H), 6.58 (d,J=16.6 Hz, 1H), 3.00 (t, J=7.6 Hz, 2H), 2.72 (t, J=7.6 Hz, 2H), 2.37 (s,6H);

cis-isomer: ¹H NMR (500 MHz, CDCl₃) δ 7.39-6.80 (m, 7H), 6.62 (d, J=12.2Hz, 1H), 6.54 (d, J=12.2 Hz, 1H), 2.73 (t, J=7.7 Hz, 2H), 2.46 (t, J=7.7Hz, 2H), 2.14 (s, 6H); ESI MS m/z 263 [M+H−H₂O]⁺.

Step 4:

To a stirred solution of 10 (0.300 g, 1.07 mmol) in DMF (10 mL) wasadded N,N-diisopropylethylamine (0.690 g, 5.34 mmol),1-hydroxybenzotriazole (0.289 g, 2.14 mmol),1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.410 g,2.14 mmol) and morpholine (0.189 g, 2.17 mmol) at room temperature.After 18 h the reaction mixture was diluted with ethyl acetate (100 mL)and washed sequentially with 10% aqueous potassium carbonate (2×20 mL),water (2×20 mL) and brine (20 mL). The organic layer was dried (Na₂SO₄),filtered and concentrated. The residue was purified by flash columnchromatography (silica gel, 60:40 hexanes/ethyl acetate) to give 11 as acolorless oil. Yield (0.335 g, 90%.), trans-/cis-isomer ratio 2:1 trans:cis.

trans-isomer: ¹H NMR (500 MHz, CDCl₃) δ 7.38-6.79 (m, 8H), 6.58 (d,J=16.6 Hz, 1H), 3.63-3.28 (m, 8H), 3.02 (t, J=7.6 Hz, 2H), 2.65 (t,J=7.6 Hz, 2H), 2.37 (s, 6H); cis-isomer: ¹H NMR (500 MHz, CDCl₃) δ7.38-6.79 (m, 7H), 6.63 (d, J=12.2 Hz, 1H), 6.53 (d, J=12.2 Hz, 1H),3.63-3.28 (m, 8H), 2.77 (t, J=7.7 Hz, 2H), 2.35 (t, J=7.7 Hz, 2H), 2.15(s, 6H); ESI MS m/z 350 [M+H]⁺.

Step 5:

To a stirred solution of 11 (0.332 g, 0.950 mmol) in THF (5 mL) wasadded lithium aluminum hydride (0.108 g, 2.85 mmol) at room temperature.After 18 h the reaction mixture was cooled to 0° C., quenched with 2Naqueous sodium hydroxide (0.23 mL), the resulting suspension was dilutedwith MTBE (50 mL), filtered and concentrated. The resulting residue waspurified by Preparative HPLC (Method 1) to give(E)-4-(3-(3-(2,6-Dimethylstyryl)phenyl)propyl)morpholine as a colorlessoil. Yield (0.114 g, 36%): R_(f) 0.81 (silica gel, 50:40:10 ethylacetate/hexanes/7N ammonia in methanol); ¹H NMR (500 MHz, CD₃OD) δ 7.34(m, 2H), 7.26 (t, J=7.5 Hz, 1H), 7.15 (d, J=16.7 Hz, 1H), 7.11 (d, J=7.5Hz, 1H), 7.03 (s, 3H), 6.56 (d, J=16.7 Hz, 1H), 3.68 (t, J=4.6 Hz, 4H),2.66 (t, J=7.5 Hz, 2H), 2.45 (br s, 4H), 2.39 (t, J=7.7 Hz, 2H), 2.33(s, 6H), 1.86 (m, 2H); ¹³C NMR (125 MHz, CD₃OD) δ 143.8, 139.2, 138.4,137.2, 135.5, 129.9, 129.0, 128.9, 128.0, 127.9, 127.6, 125.0, 67.8,59.6, 54.9, 34.7, 29.2, 21.3; ESI MS m/z 336 [M+H]⁺; HPLC (Method 5)>99%(AUC), t_(R)=13.86 min. HRMS Calcd for C₂₃H₂₉NO [M+H]: 336.2327. Found:336.2312.

Example 4 Preparation of(Z)-4-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)morpholine

(Z)-4-(3-(3-(2,6-Dimethylstyryl)phenyl)propyl)morpholine was isolatedduring the synthesis of Example 3 as a colorless oil. Yield (0.039 g,12%): ¹H NMR (500 MHz, CD₃OD) δ 7.05 (m, 4H), 6.95 (d, J=7.6 Hz, 1H),6.83 (d, J=7.7 Hz, 1H), 6.77 (s, 1H), 6.66 (d, J=12.2 Hz, 1H), 6.54 (d,J=12.2 Hz, 1H), 3.68 (t, J=4.6 Hz, 4H), 2.38 (m, 6H), 2.21 (t, J=7.5 Hz,2H), 2.13 (s, 6H), 1.58 (m, 2H); ¹³C NMR (125 MHz, CD₃OD) δ 143.1,139.1, 138.7, 136.7, 132.5, 129.8, 129.4, 129.0, 128.7, 128.6, 128.2,127.3, 67.8, 59.5, 54.9, 34.5, 28.8, 20.6; ESI MS m/z 336 [M+H]⁺. HPLC(Method 5)>99% (AUC), t_(R)=14.40 min. HRMS Calcd for C₂₃H₂₉NO [M+H]:336.2327. Found: 336.2311.

Example 5 Preparation of(E)-1-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)pyrrolidine

(E)-1-(3-(3-(2,6-Dimethylstyryl)phenyl)propyl)pyrrolidine was preparedfollowing the method used in Example 3.

Step 1:

Coupling of acid 10 with pyrrolidine gave3-(3-(2,6-dimethylstyryl)phenyl)-1-(pyrrolidin-1-yl)propan-1-one as acolorless oil. Yield (0.26 g, 90%), isomer ratio 2:1 trans: cis.

trans-isomer: ¹H NMR (500 MHz, CDCl₃) δ 7.36-6.82 (m, 8H), 6.58 (d,J=16.6 Hz, 1H), 3.48 (t, J=6.9 Hz, 2H), 3.31 (t, J=6.9 Hz, 2H), 3.02 (t,J=7.6 Hz, 2H), 2.60 (t, J=7.6 Hz, 2H), 2.37 (s, 6H), 1.92-1.79 (m, 4H);

cis-isomer: ¹H NMR (500 MHz, CDCl₃) δ 7.36-6.82 (m, 7H), 6.63 (d, J=12.2Hz, 1H), 6.52 (d, J=12.2 Hz, 1H), 3.44 (t, J=6.9 Hz, 2H), 3.22 (t, J=6.9Hz, 2H), 2.78 (t, J=7.7 Hz, 2H), 2.31 (t, J=7.7 Hz, 2H), 2.15 (s, 6H),1.92- 1.79 (m, 4H); ESI MS m/z 334 [M+H]⁺.

Step 2:

Reduction of3-(3-(2,6-dimethylstyryl)phenyl)-1-(pyrrolidin-1-yl)propan-1-onefollowed by purification by Preparative HPLC (Method 1) gave Example 5(0.095 g, 39%) as a colorless oil: R_(f) 0.75 (silica gel, 50:40:10ethyl acetate/hexanes/7N ammonia in methanol); ¹H NMR (500 MHz, CD₃OD) δ7.34 (m, 2H), 7.27 (t, J=7.5 Hz, 1H), 7.16 (d, J=16.7 Hz, 1H), 7.11 (d,J=7.5 Hz, 1H), 7.03 (s, 3H), 6.57 (d, J=16.7 Hz, 1H), 2.67 (t, J=7.6 Hz,2H), 2.53 (m, 6H), 2.34 (s, 6H), 1.88 (m, 2H), 1.80 (m, 4H); ¹³C NMR(125 MHz, CD₃OD) δ 143.8, 139.2, 138.4, 137.3, 135.5, 129.9, 129.0,128.9, 128.0, 127.8, 127.6, 125.0, 57.2, 55.2, 34.9, 31.6, 24.3, 21.3;ESI MS m/z 320 [M+H]⁺. HPLC (Method 5)>99% (AUC), t_(R)=15.21 min. HRMSCalcd for C₂₃H₂₉N [M+H]: 320.2378. Found: 320.2376.

Example 6 Preparation of(Z)-1-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)pyrrolidine

(Z)-1-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)pyrrolidine was isolatedduring the synthesis of Example 5 as a colorless oil. Yield (0.035 g,14%); R_(f) 0.75 (silica gel, 50:40:10 ethyl acetate/hexanes/7N ammoniain methanol); ¹H NMR (500 MHz, CD₃OD) δ 7.05 (m, 4H), 6.95 (d, J=7.6 Hz,1H), 6.83 (d, J=7.7 Hz, 1H), 6.77 (s, 1H), 6.65 (d, J=12.2 Hz, 1H), 6.54(d, J=12.2 Hz, 1H), 2.48 (m, 4H), 2.39 (t, J=7.6 Hz, 2H), 2.33 (m, 2H),2.13 (s, 6H), 1.79 (m, 4H), 1.61 (m, 2H); ¹³C NMR (125 MHz, CD₃OD) δ143.1, 139.1, 138.7, 136.7, 132.5, 129.8, 129.4, 129.0, 128.7, 128.6,128.2, 127.3, 57.1, 55.2, 34.8, 31.3, 24.3, 20.6; ESI MS m/z 320 [M+H]⁺.HPLC (Method 5)>99% (AUC), t_(R)=15.56 min. HRMS Calcd for C₂₃H₂₉N[M+H]: 320.2378. Found: 320.2365.

Example 7 Preparation of(E)-3-(3-(2,6-dimethylstyryl)phenyl)-N-methylpropan-1-amine

(E)-3-(3-(2,6-Dimethylstyryl)phenyl)-N-methylpropan-1-amine was preparedfollowing the method used to prepare Example 3.

Step 1:

Coupling of acid 10 with methylamine gave3-(3-(2,6-dimethylstyryl)phenyl)-N-methylpropanamide as a colorless oil.Yield (0.277 g, 88%), isomer ratio 2:1 trans: cis.

trans-isomer: ¹H NMR (300 MHz, CDCl₃) δ 7.37-6.78 (m, 8H), 6.57 (d,J=16.6 Hz, 1H), 5.47 (br s, 1H), 3.00 (t, J=7.4 Hz, 2H), 2.78 (d, J=4.8Hz, 3H), 2.49 (t, J=7.4 Hz, 2H), 2.36 (s, 6H);

cis-isomer: ¹H NMR (500 MHz, CDCl₃) δ 7.37-6.78 (m, 7H), 6.63 (d, J=12.2Hz, 1H), 6.53 (d, J=12.2 Hz, 1H), 5.18 (br s, 1H), 2.76 (t, J=7.4 Hz,2H), 2.71 (d, J=4.8 Hz, 3H), 2.17 (t, J=7.4 Hz, 2H), 2.14 (s, 6H).

Step 2:

Reduction of 3-(3-(2,6-dimethylstyryl)phenyl)-N-methylpropanamidefollowed by purification by Preparative HPLC (Method 1) gave Example 7(0.080 g, 30%) as a colorless oil: R_(f) 0.65 (silica gel, 50:40:10ethyl acetate/hexanes/7N ammonia in methanol); ¹H NMR (500 MHz, CD₃OD) δ7.35 (m, 2H), 7.27 (t, J=7.5 Hz, 1H), 7.16 (d, J=16.7 Hz, 1H), 7.11 (d,J=7.5 Hz, 1H), 7.03 (s, 3H), 6.57 (d, J=16.7 Hz, 1H), 2.67 (t, J=7.6 Hz,2H), 2.58 (t, J=7.6 Hz, 2H), 2.36 (s, 3H), 2.34 (s, 6H), 1.85 (m, 2H);¹³C NMR (75 MHz, CD₃OD) δ 143.8, 139.2, 138.4, 137.3, 135.5, 129.9,129.0, 128.9, 128.0, 127.9, 127.5, 125.0, 52.3, 36.2, 34.6, 32.2, 21.3;ESI MS m/z 280 [M+H]⁺; HPLC (Method 5) 90.4% (AUC), t_(R)=14.39 min.HRMS Calcd for C₂₀H₂₅N [M+H]: 280.2065. Found: 280.2062.

Example 8 Preparation of(Z)-3-(3-(2,6-dimethylstyryl)phenyl)-N-methylpropan-1-amine

(Z)-3-(3-(2,6-Dimethylstyryl)phenyl)-N-methylpropan-1-amine was isolatedduring the synthesis of Example 7 as a colorless oil. Yield (0.025 g,10%): R_(f) 0.65 (silica gel, 50:40:10 ethyl acetate/hexanes/7N ammoniain methanol); ¹H NMR (500 MHz, CD₃OD) δ 7.05 (m, 4H), 6.96 (d, J=7.6 Hz,1H), 6.82 (d, J=7.7 Hz, 1H), 6.78 (s, 1H), 6.65 (d, J=12.2 Hz, 1H), 6.54(d, J=12.2 Hz, 1H), 2.40 (m, 4H), 2.32 (s, 3H), 2.12 (s, 6H), 1.60 (m,2H); ¹³C NMR (75 MHz, CD₃OD) δ 143.0, 139.1, 138.7, 136.7, 132.5, 129.9,129.4, 129.1, 128.7, 128.6, 128.2, 127.1, 60.0, 36.0, 34.4, 31.6, 20.5;ESI MS m/z 280 [M+H]⁺; HPLC (Method 5) 97.3% (AUC), t_(R)=14.64 min.HRMS Calcd for C₂₀H₂₅N [M+H]: 280.2065. Found: 280.2063.

Example 9 Preparation of(E)-3-(3-(2,6-dimethylstyryl)phenyl)-N,N-dimethylpropan-1-amine

(E)-3-(3-(2,6-Dimethylstyryl)phenyl)-N,N-dimethylpropan-1-amine wasprepared following the method used to prepare Example 3.

Step 1:

Coupling of acid 10 with dimethylamine gave(E)-3-(3-(2,6-dimethylstyryl)phenyl)-N,N-dimethylpropanamide as acolorless oil. Yield (0.273 g, 83%), isomer ratio 2:1 trans: cis.

trans-isomer: ¹H NMR (300 MHz, CDCl₃) δ 7.38-6.81 (m, 8H), 6.58 (d,J=16.6 Hz, 1H), 3.03-2.87 (m, 8H), 2.65 (t, J=7.6 Hz, 2H), 2.37 (s, 6H);

cis-isomer: ¹H NMR (500 MHz, CDCl₃) δ 7.38-6.81 (m, 7H), 6.63 (d, J=12.2Hz, 1H), 6.53 (d, J=12.2 Hz, 1H), 3.03-2.87 (m, 6H), 2.76 (t, J=7.7 Hz,2H), 2.36 (t, J=7.7 Hz, 2H), 2.15 (s, 6H).

Step 2:

Reduction of 3-(3-(2,6-dimethylstyryl)phenyl)-N,N-dimethylpropanamidefollowed by purification by Preparative HPLC (Method 1) gave Example 9(0.101 g, 39%) as a colorless oil: R_(f) 0.81 (silica gel, 50:40:10ethyl acetate/hexanes/7N ammonia in methanol); ¹H NMR (500 MHz, CD₃OD) δ7.35 (m, 2H), 7.27 (t, J=7.5 Hz, 1H), 7.16 (d, J=16.7 Hz, 1H), 7.11 (d,J=7.5 Hz, 1H), 7.03 (s, 3H), 6.57 (d, J=16.7 Hz, 1H), 2.65 (t, J=7.6 Hz,2H), 2.37 (m, 2H), 2.34 (s, 6H), 2.24 (s, 6H), 1.84 (m, 2H); ¹³C NMR(125 MHz, CD₃OD) δ 143.7, 139.2, 138.4, 137.3, 135.5, 129.9, 129.0,128.9, 128.0, 127.8, 127.6, 125.0, 60.3, 45.6, 34.7, 30.3, 21.3; ESI MSm/z 294 [M+H]⁺; HPLC (Method 5)>99% (AUC), t_(R)=15.33 min. HRMS Calcdfor C₂₁H₂₇N [M+H]: 294.2222. Found: 294.2219.

Example 10 Preparation of(Z)-3-(3-(2,6-dimethylstyryl)phenyl)-N,N-dimethylpropan-1-amine

(Z)-3-(3-(2,6-Dimethylstyryl)phenyl)-N,N-dimethylpropan-1-amine wasisolated during the synthesis of Example 9 as a colorless oil. Yield(0.038 g, 15%): R_(f) 0.81 (silica gel, 50:40:10 ethylacetate/hexanes/7N ammonia in methanol); ¹H NMR (500 MHz, CD₃OD) δ 7.05(m, 4H), 6.95 (d, J=7.6 Hz, 1H), 6.83 (d, J=7.7 Hz, 1H), 6.77 (s, 1H),6.65 (d, J=12.2 Hz, 1H), 6.54 (d, J=12.2 Hz, 1H), 2.38 (t, J=7.6 Hz,2H), 2.18 (m, 8H), 2.12 (s, 6H), 1.55 (m, 2H); ¹³C NMR (125 MHz, CD₃OD)δ 143.0, 139.1, 138.7, 136.7, 132.4, 129.8, 129.4, 129.0, 128.7, 128.6,128.2, 127.3, 60.2, 45.5, 34.6, 29.9, 20.6; ESI MS m/z 294 [M+H]⁺; HPLC(Method 5)>99% (AUC), t_(R)=15.49 min. HRMS Calcd for C₂₁H₂₇N [M+H]:294.2222. Found: 294.2224.

Example 11 Preparation of(E)-1-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)piperidine

(E)-1-(3-(3-(2,6-Dimethylstyryl)phenyl)propyl)piperidine was preparedfollowing the method used to prepare Example 3.

Step 1:

Coupling of acid 10 with piperidine gave3-(3-(2,6-dimethylstyryl)phenyl)-1-(piperidin-1-yl)propan-1-one as acolorless oil. Yield (0.330 g, 89%), isomer ratio 2:1 trans: cis.

trans-isomer: ¹H NMR (300 MHz, CDCl₃) δ 7.37-6.80 (m, 8H), 6.58 (d,J=16.6 Hz, 1H), 3.57 (t, J=5.7 Hz, 2H), 3.36 (t, J=5.5 Hz, 2H), 3.00 (t,J=7.5 Hz, 2H), 2.65 (t, J=7.5 Hz, 2H), 2.37 (s, 6H), 1.64-1.47 (m, 6H);

cis-isomer: ¹H NMR (500 MHz, CDCl₃) δ 7.37-6.80 (m, 7H), 6.63 (d, J=12.2Hz, 1H), 6.53 (d, J=12.2 Hz, 1H), 3.53 (t, J=5.7 Hz, 2H), 3.26 (t, J=5.5Hz, 2H), 2.75 (t, J=7.5 Hz, 2H), 2.35 (t, J=7.5 Hz, 2H), 2.15 (s, 6H),1.64- 1.47 (m, 6H); ESI MS m/z 348 [M+H]⁺.

Step 2:

Reduction of3-(3-(2,6-dimethylstyryl)phenyl)-1-(piperidin-1-yl)propan-1-one followedby purification by Preparative HPLC (Method 1) gave Example 11 (0.106 g,34%) as a colorless oil: R_(f) 0.73 (silica gel, 50:40:10 ethylacetate/hexanes/7N ammonia in methanol); ¹H NMR (500 MHz, CD₃OD) δ 7.34(m, 2H), 7.26 (t, J=7.5 Hz, 1H), 7.16 (d, J=16.7 Hz, 1H), 7.10 (d, J=7.5Hz, 1H), 7.03 (s, 3H), 6.56 (d, J=16.7 Hz, 1H), 2.64 (t, J=7.6 Hz, 2H),2.38 (m, 6H), 2.34 (s, 6H), 1.87 (m, 2H), 1.60 (m, 4H), 1.46 (m, 2H);¹³C NMR (75 MHz, CD₃OD) δ 143.8, 139.2, 138.4, 137.3, 135.5, 129.9,129.0, 128.9, 128.0, 127.9, 127.6, 125.0, 60.1, 55.7, 34.9, 29.4, 26.6,25.4, 21.3; ESI MS m/z 334 [M+H]⁺. HPLC (Method 5)>99% (AUC),t_(R)=13.14 min. HRMS Calcd for C₂₄H₃₁N [M+H]: 334.2535. Found:334.2520.

Example 12 Preparation of(Z)-1-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)piperidine

(Z)-1-(3-(3-(2,6-Dimethylstyryl)phenyl)propyl)piperidine was isolatedduring the synthesis of Example 11 as a colorless oil. Yield (0.027 g,9%): R_(f) 0.73 (silica gel, 50:40:10 ethyl acetate/hexanes/7N ammoniain methanol); ¹H NMR (500 MHz, CD₃OD) δ 7.05 (m, 4H), 6.95 (d, J=7.6 Hz,1H), 6.83 (d, J=7.7 Hz, 1H), 6.76 (s, 1H), 6.65 (d, J=12.2 Hz, 1H), 6.54(d, J=12.2 Hz, 1H), 2.36 (m, 6H), 2.19 (m, 2H), 2.12 (s, 6H), 1.58 (m,6H), 1.46 (m, 2H); ¹³C NMR (75 MHz, CD₃OD) δ 143.1, 139.0, 138.7, 136.7,132.5, 129.8, 129.4, 128.9, 128.7, 128.6, 128.1, 127.3, 59.9, 55.6,34.8, 28.9, 26.6, 25.3, 20.6; ESI MS m/z 334 [M+H]⁺; HPLC (Method 5)98.7% (AUC), t_(R)=13.72 min. HRMS Calcd for C₂₄H₃₁N [M+H]: 334.2535.Found: 334.2527.

Example 13 Preparation of(e)-1-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)piperazine

(E)-3-(3-(2,6-dimethylstyryl)phenyl)-1-(piperazin-1-yl)propan-1-one wasprepared following the method shown in Scheme 3.

Step 1:

tert-Butyl4-(3-(3-(2,6-Dimethylstyryl)phenyl)propanoyl)piperazine-1-carboxylate(12) was prepared from Compound 10 and N-Boc-piperazine following themethod used to prepare Example 3. Yield (0.445 g, 95%) of a colorlessoil as a mixture of trans-/cis-isomers. Isomer ratio 2:1 trans: cis.

trans-isomer: ¹H NMR (300 MHz, CDCl₃) δ 7.38-6.79 (m, 8H), 6.57 (d,J=16.6 Hz, 1H), 3.62-3.26 (m, 8H), 3.02 (t, J=7.5 Hz, 2H), 2.67 (t,J=7.5 Hz, 2H), 2.36 (s, 6H), 1.46 (s, 9H);

cis-isomer: ¹H NMR (500 MHz, CDCl₃) δ 7.38-6.79 (m, 7H), 6.63 (d, J=12.2Hz, 1H), 6.53 (d, J=12.2 Hz, 1H), 3.62-3.26 (m, 8H), 2.76 (t, J=7.5 Hz,2H), 2.35 (t, J=7.5 Hz, 2H), 2.15 (s, 6H), 1.48 (s, 9H); ESI MS m/z 449[M+H]⁺.

Step 2:

To a stirred solution of tert-Butyl4-(3-(3-(2,6-Dimethylstyryl)phenyl)propanoyl)piperazine-1-carboxylate(12) (0.450 g, 1.00 mmol) in methylene chloride (6 mL) was addedtrifluoroacetic acid (3 mL) at room temperature. After 1 h, the reactionmixture was basified by slow addition of saturated aqueous sodiumbicarbonate (30 mL) followed by 10% aqueous potassium carbonate to pH10. The resulting mixture was extracted with hexanes (3×50 mL) and thecombined extracts were dried (Na₂SO₄), filtered and concentrated. Theresulting residue was purified by Preparative HPLC (Method 1) to give(E)-3-(3-(2,6-dimethylstyryl)phenyl)-1-(piperazin-1-yl)propan-1-one (13)(0.147 g, 42%) as a colorless oil. R_(f) 0.58 (silica gel, 50:40:10ethyl acetate/hexanes/7N ammonia in methanol); ¹H NMR (500 MHz, CD₃OD) δ7.41 (s, 1H), 7.37 (d, J=7.7 Hz, 1H), 7.28 (t, J=7.5 Hz, 1H), 7.18 (d,J=16.7 Hz, 1H), 7.15 (d, J=7.5 Hz, 1H), 7.03 (s, 3H), 6.57 (d, J=16.7Hz, 1H), 3.53 (t, J=5.1 Hz, 2H), 3.41 (t, J=5.1 Hz, 2H), 2.95 (t, J=7.4Hz, 2H), 2.71 (m, 4H), 2.59 (t, J=5.1 Hz, 2H), 2.34 (s, 6H); ¹³C NMR (75MHz, CD₃OD) δ 173.5, 142.8, 139.3, 138.3, 137.3, 135.4, 130.1, 129.1,129.0, 128.3, 127.9, 127.7, 125.5, 47.9, 46.7, 46.4, 43.6, 35.5, 32.9,21.3; ESI MS m/z 349 [M+H]⁺. HPLC (Method 5)>99% (AUC), t_(R)=10.63 min.HRMS Calcd for C₂₃H₂₈N₂O [M+H]: 349.2280. Found: 349.2275.

Step 3:

To a stirred solution of(E)-3-(3-(2,6-dimethylstyryl)phenyl)-1-(piperazin-1-yl)propan-1-one (13)(0.138 g, 0.396 mmol) in THF (5 mL) was added borane-THF complex (0.79mL, 1M solution in THF, 0.790 mmol) and the reaction mixture heated toreflux. After 1 h the reaction mixture was cooled to room temperatureand another portion of borane-THF complex (0.40 mL, 1M solution in THF,0.400 mmol) was added. After 18 h the reaction was quenched by slowaddition of methanol (10 mL), concentrated, the resulting residuesuspended in 6N hydrochloric acid (5 mL) and heated at 90° C. After 0.5h the mixture was cooled to room temperature and basified to pH 12 with2N aqueous sodium hydroxide. The resulting suspension was extracted withmethylene chloride (3×50 mL) and the combined extracts dried (Na₂SO₄),filtered and concentrated. The resulting residue was purified by flashcolumn chromatography (silica gel, 50:46:4 ethyl acetate/hexanes/7Nammonia in methanol) to give Example 13 as a colorless oil. Yield (0.062g, 47%): R_(f) 0.60 (silica gel, 50:40:10 ethyl acetate/hexanes/7Nammonia in methanol); ¹H NMR (500 MHz, CD₃OD) δ 7.35 (m, 2H), 7.27 (t,J=7.5 Hz, 1H), 7.17 (d, J=16.7 Hz, 1H), 7.11 (d, J=7.5 Hz, 1H), 7.03 (s,3H), 6.57 (d, J=16.7 Hz, 1H), 2.84 (t, J=5.0 Hz, 4H), 2.66 (t, J=7.6 Hz,2H), 2.41 (m, 6H), 2.34 (s, 6H), 1.86 (m, 2H); ¹³C NMR (125 MHz, CD₃OD)δ 143.8, 139.2, 138.4, 137.2, 135.5, 129.9, 129.0, 128.9, 128.0, 127.9,127.6, 125.0, 59.8, 54.9, 46.2, 34.7, 29.2, 21.3; ESI MS m/z 335 [M+H]⁺;HPLC (Method 5)>99% (AUC), t_(R)=10.11 min. HRMS Calcd for C₂₃H₃₀N₂[M+H]: 335.2487. Found: 335.2491.

Example 14 Preparation of(Z)-1-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)piperazine

(Z)-1-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)piperazine was preparedfollowing the method used to prepare Example 3.

Step 1:

(Z)-3-(3-(2,6-dimethylstyryl)phenyl)-1-(piperazin-1-yl)propan-1-one wasisolated during the synthesis of Example 13 as a colorless oil. Yield(0.073 g, 21%): R_(f)0.58 (silica gel, 50:40:10 ethyl acetate/hexanes/7Nammonia in methanol); ¹H NMR (500 MHz, CD₃OD) δ 7.05 (m, 4H), 6.98 (d,J=7.6 Hz, 1H), 6.84 (d, J=7.7 Hz, 1H), 6.80 (s, 1H), 6.66 (d, J=12.2 Hz,1H), 6.55 (d, J=12.2 Hz, 1H), 3.50 (t, J=5.0 Hz, 2H), 3.31 (m, 2H), 2.68(m, 6H), 2.40 (t, J=8.2 Hz, 2H), 2.12 (s, 6H); ¹³C NMR (75 MHz, CD₃OD) δ173.2, 142.1, 139.2, 138.7, 136.8, 132.3, 130.0, 129.6, 129.1, 128.7,128.2, 127.4, 44.8, 46.8, 46.4, 43.6, 35.4, 32.6, 20.6; ESI MS m/z 349[M+H]⁺; HPLC (Method 5)>99% (AUC), t_(R)=11.12 min. HRMS Calcd forC₂₃H₂₈N₂O [M+H]: 349.2280. Found: 349.2271.

Step 2:

(Z)-1-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)piperazine was preparedfollowing the method used in Example 13. Yield (0.046 g, 75%) of acolorless oil: R_(f) 0.60 (silica gel, 50:40:10 ethyl acetate/hexanes/7Nammonia in methanol); ¹H NMR (500 MHz, CD₃OD) δ 7.05 (m, 4H), 6.95 (d,J=7.6 Hz, 1H), 6.83 (d, J=7.7 Hz, 1H), 6.76 (s, 1H), 6.65 (d, J=12.2 Hz,1H), 6.54 (d, J=12.2 Hz, 1H), 2.83 (t, J=5.0 Hz, 4H), 2.38 (m, 6H), 2.20(m, 2H), 2.12 (s, 6H), 1.58 (m, 2H); ¹³C NMR (75 MHz, CD₃OD) δ 143.1,139.1, 138.7, 136.7, 132.5, 129.8, 129.4, 129.0, 128.7, 128.6, 128.1,127.3, 59.7, 54.9, 46.2, 34.6, 28.9, 20.6; ESI MS m/z 335 [M+H]⁺; HPLC(Method 5) 97.0% (AUC), t_(R)=9.92 min. HRMS Calcd for C₂₃H₃₀N₂ [M+H]⁺:335.2487. Found: 335.2491.

Example 15 Preparation of(E)-2-amino-N-(3-(2,6-dimethylstyryl)phenyl)acetamide

(E)-3-amino-N-(3-(2,6-dimethylstyryl)phenyl)acetamide was preparedfollowing the method described in Scheme 4.

Step 1:

Coupling of Wittig reagent 3 with 3-nitrobenzaldehyde following themethod used to prepare Example 4 gave olefin 14 as a yellow solid. Yield(0.486 g, 87%), isomer ratio 3:1 trans: cis.

cis-isomer: ¹H NMR (500 MHz, CDCl₃) δ 7.96 (d, J=7.2 Hz, 1H), 7.87 (d,J=1.6 Hz, 1H), 7.25-7.23 (m, 1H), 7.16-7.06 (m, 4H), 6.74 (d, J=11.4 Hz,1H), 6.69 (d, J=12.2 Hz, 1H), 2.14 (s, 6H);

trans-isomer: ¹H NMR (500 MHz, CDCl₃) δ 8.34 (d, J=1.9 Hz, 1H),8.12-8.10 (m, 1H), 7.78 (d, J=7.8 Hz, 1H), 7.53 (t, J=8.0 Hz, 1H),7.25-7.23 (m, 1H), 7.16-7.06 (m, 3H), 6.65 (d, J=16.6 Hz, 1H), 2.38 (s,6H).

Step 2:

To a stirred solution of 14 (0.486 g, 1.9 mmol) in ethanol (25 mL) wasadded iron (1.08 g, 10 mmol) and 1N HCl (2.0 mL, 2.0 mmol) and theresulting suspension was heated at 60° C. After 1.5 h the mixture wascooled to room temperature and filtered over diatomaceous earth. Thefilter cake was washed with ethanol (50 mL), the filtrate concentrated,the residue was dissolved in ethyl acetate (100 mL), and the resultingsolution washed with saturated aqueous sodium bicarbonate (2×50 mL). Thecombined aqueous phases were extracted with ethyl acetate (75 mL) andthe combined organics were dried (MgSO₄), filtered and concentrated.Purification by column chromatography (silica, 0-15% ethylacetate/hexanes) provided (0.071 g, 17%) of 15 as a yellow solid and(0.348 g, 82%) of 16 as a yellow solid.

15: ¹H NMR (500 MHz, CDCl₃) δ 7.10 (t, J=7.7 Hz, 1H), 7.02 (d, J=7.5 Hz,2H), 6.92 (t, J=7.8 Hz, 1H), 6.55 (d, J=12.3 Hz, 1H), 6.49 (d, J=12.5Hz, 1H), 6.47-6.45 (m, 1H), 6.41 (d, J=7.6 Hz, 1H), 6.31-6.30 (m, 1H),3.43 (br s, 2H), 2.16 (s, 6H).

16: ¹H NMR (500 MHz, CDCl₃) δ 7.16 (t, J=7.8 Hz, 1H), 7.08-7.04 (m, 4H),6.91 (d, J=7.8 Hz, 1H), 6.84-6.83 (m, 1H), 7.62 (dd, J=7.9, 2.2 Hz, 1H),5.52 (d, J=16.7 Hz, 1H), 3.69 (br s, 2H), 2.36 (s, 6H).

Step 3:

To a stirred solution of 16 (0.132 g, 0.590 mmol) and Fmoc-β-alanine(0.306 g, 0.980 mmol) in THF (5 mL) was added HBTU (0.490 g, 1.31 mmol),followed by N,N-diisopropylethylamine (0.245 g, 1.89 mmol) The reactionmixture was stirred overnight, then concentrated under reduced pressure.The residue was purified by flash chromatography (silica gel, 0-50%ethyl acetate/hexanes) followed by trituration with methylene chlorideto provide 17 as a white solid. Yield (0.163 g, 53%): ¹H NMR (500 MHz,CDCl₃) δ 7.74 (d, J=7.4 Hz, 2H), 7.69 (s, 1H), 7.57 (d, J=7.3 Hz, 2H),7.45-7.27 (m, 8H), 7.12 (d, J=16.5 Hz, 1H), 7.10-7.06 (m, 3H), 6.60 (d,J=16.7 Hz, 1H), 5.47 (br s, 1H), 4.39 (d, J=6.6 Hz, 2H), 4.21 (d, J=6.8Hz, 1H), 3.59 (d, J=4.9 Hz, 2H), 2.65 (br s, 2H), 2.36 (s, 6H)); ESI MSm/z 517 [M+H]⁺.

Step 4:

To a stirred solution of 16 (0.135 g, 0.600 mmol) and Fmoc-glycine(0.294 g, 0.990 mmol) in THF (5 mL) was added HBTU (0.494 g, 1.31 mmol),followed by N,N-diisopropylethylamine (0.245 g, 1.89 mmol). The reactionmixture was stirred overnight, then concentrated under reduced pressure.The residue was purified by flash chromatography (silica gel, 0-50%ethyl acetate/hexanes) to provide 17 as a white solid. Yield (0.311 g,quant.): ¹H NMR (500 MHz, CDCl₃) δ 8.10 (br s, 1H), 7.34 (d, J=7.5 Hz,2H), 7.65 (s, 1H), 7.57 (d, J=7.3 Hz, 2H), 7.39-7.36 (m, 3H), 7.31-7.27(m, 4H), 7.12-7.05 (m, 4H), 6.54 (d, J=16.6 Hz, 1H), 5.64 (br s, 1H),4.48 (d, J=6.8 Hz, 2H), 4.23 (t, J=6.8 Hz, 1H), 4.03 (br s, 2H), 2.35(s, 6H); ESI MS m/z 503 [M+H]⁺.

Step 5:

A solution of 17 (0.311 g, 0.60 mmol) in diethylamine (5 mL) was stirredat room temperature. After 4 h the reaction mixture was concentrated toan oily residue which was purified by flash column chromatography(silica gel, 1-10% 7N ammonia in methanol/methylene chloride) to provideExample 15 as a yellow gum. Yield (0.119 g, 71%): R_(f) 0.59 (silicagel, 95:5 methylene chloride/7N ammonia in methanol); ¹H NMR (500 MHz,CD₃OD) δ 7.81 (s, 1H), 7.47 (d, J=7.8 Hz, 1H), 7.31 (t, J=7.8 Hz, 1H),7.27 (d, J=7.7 Hz, 1H), 7.19 (d, J=16.7 Hz, 1H), 7.04 (s, 3H), 6.57 (d,J=16.6 Hz, 1H), 3.43 (s, 2H), 2.35 (s, 6H); ¹³C NMR (125 MHz, CD₃OD) δ173.6, 140.0, 139.7, 138.0, 137.1, 134.9, 130.2, 128.9, 128.6, 127.8,123.3, 120.2, 118.6, 45.7, 21.2; ESI MS m/z 281 [M+H]⁺; HPLC (Method5)>99% (AUC), t_(R)=10.88 min. HRMS calcd for C₁₈H₂₀N₂O [M+H]: 281.1654.Found: 281.1664.

Example 16 Preparation of (E)-3-(3-(2-methylstyryl)phenyl)propan-1-amine

(E)-3-(3-(2-methylstyryl)phenyl)propan-1-amine was prepared followingthe method used in Example 1.

Step 1:

Coupling of 3-iodobenzaldehyde with (2-methylbenzyl)triphenylphosphoniumbromide followed by flash chromatography (silica gel, eluent: hexanes)gave (E)-1-(3-iodostyryl)-2-methylbenzene as a colorless oil. Yield(0.498 g, 36%): ¹H NMR (500 MHz, CDCl₃) δ 7.87-7.86 (m, 1H), 7.59-7.55(m, 2H), 7.45 (d, J=7.8 Hz, 1H), 7.30 (d, J=16.2 Hz, 1H), 7.23-7.18 (m,3H), 7.09 (t, J=7.9 Hz, 1H), 6.86 (d, J=16.2 Hz, 1H), 2.43 (s, 3H) and(Z)-1-(3-iodostyryl)-2-methylbenzene as a colorless oil. Yield (0.0.540g, 39%): ¹H NMR (500 MHz, CDCl₃) δ 7.47-7.45 (m, 2H), 7.21-7.15 (m, 2H),7.11-7.01 (m, 3H), 6.85 (t, J=7.8 Hz, 1H), 6.68 (d, J=12.2 Hz, 1H), 6.49(d, J=12.1 Hz, 1H), 2.26 (s, 3H).

Step 2:

Coupling of (E)-1-(3-iodostyryl)-2-methylbenzene with allyl alcohol gave(E)-3-(3-(2-Methylstyryl)phenyl)propanal as a yellow oil. Yield (0.103g, 52%): ¹H NMR (500 MHz, CDCl₃) δ 9.85 (t, J=1.2 Hz, 1H), 7.59 (d,J=7.2 Hz, 1H), 7.38 (d, J=7.8 Hz, 1H), 7.33-7.28 (m, 3H), 7.22-7.18 (m,3H), 7.10 (d, J=7.5 Hz, 1H), 6.96 (d, J=16.1 Hz, 1H), 2.99 (t, J=7.6 Hz,2H), 2.84-2.31 (m, 2H), 2.44 (s, 3H); ESI MS m/z 233 [M+H−H₂O]]⁺.

Step 3:

Reductive amination of (E)-3-(3-(2-Methylstyryl)phenyl)propanal withammonia gave Example 16 as a yellow oil. Yield (0.017 g, 16%): R_(f)0.35 (silica gel, 95:5 methylene chloride/7N ammonia in methanol); ¹HNMR (500 MHz, CD₃OD) δ 7.58 (d, J=7.3 Hz, 1H), 7.40-7.37 (m, 3H), 7.27(t, J=7.6 Hz, 1H), 7.17-7.10 (m, 4H), 7.00 (d, J=16.2 Hz, 1H), 2.73 (t,J=7.4 Hz, 2H), 2.69 (t, J=7.7 Hz, 2H), 2.41 (s, 3H), 1.86 (quint, J=7.5Hz, 2H); ¹³C NMR (75 MHz, CD₃OD) δ 143.8, 139.2, 137.7, 136.9, 131.4,131.2, 129.8, 128.8, 128.5, 127.7, 127.3, 126.3, 125.1, 42.1, 35.6,34.2, 20.0; ESI MS m/z 252 [M+H]⁺; HPLC (Method 2) 98.6% (AUC),t_(R)=8.86 min. HRMS calcd for C₁₈H₂₁N [M+H]: 252.1752. Found: 252.1748.

Example 17 Preparation of (Z)-3-(3-(2-methylstyryl)phenyl)propan-1-amine

(Z)-3-(3-(2-methylstyryl)phenyl)propan-1-amine was prepared followingthe method used in Example 1.

Step 1:

Coupling of (Z)-1-(3-iodostyryl)-2-methylbenzene with allyl alcohol gave(Z)-3-(3-(2-Methylstyryl)phenyl)propanal as a yellow oil. Yield (0.075g, 37%): ¹H NMR (500 MHz, CDCl₃) δ 9.70 (s, 1H), 7.21-7.03 (m, 5H),6.97-6.95 (m, 2H), 6.89 (s, 1H), 6.65 (d, J=12.1 Hz, 1H), 6.59 (d,J=12.1 Hz, 1H), 2.78 (t, J=7.5 Hz, 2H), 2.58-2.55 (m, 2H), 2.26 (s, 3H);ESI MS m/z 233 [M+H−H₂O]⁺.

Step 2:

Reductive amination of (Z)-3-(3-(2-Methylstyryl)phenyl)propanal withammonia gave Example 22 as a yellow oil. Yield (0.016 g, 21%): R_(f)0.57 (silica gel, 95:5 methylene chloride/7N ammonia in methanol); ¹HNMR (500 MHz, CD₃OD) δ 7.18 (d, J=7.6 Hz, 1H), 7.15-7.12 (m, 1H),7.06-7.01 (m, 3H), 6.96 (d, J=7.7 Hz, 1H), 6.89 (d, J=7.4 Hz, 2H), 6.65(d, J=12.2 Hz, 1H), 6.61 (d, J=12.2 Hz, 1H), 4.50 (t, J=7.3 Hz, 2H),2.44 (t, J=7.6 Hz, 2H), 2.22 (s, 3H), 1.59 (quint, J=7.4 Hz, 2H); ¹³CNMR (75 MHz, CD₃OD) δ142.9, 138.7, 138.5, 137.1, 131.9, 131.1, 130.5,130.0, 129.9, 129.2, 128.3, 127.6, 126.8, 41.9, 35.3, 34.0, 20.0; ESI MSm/z 252 [M+H]⁺; HPLC (Method 2) 92.9% (AUC), t_(R)=11.53 min. HRMS calcdfor C₁₈H₂₁N [M+H]: 252.1752. Found: 252.1761.

Example 18 Preparation of(E)-2-(3-(2,6-dimethylstyryl)phenylthio)ethanamine

(E)-2-(3-(2,6-dimethylstyryl)phenylthio)ethanamine was preparedfollowing the method shown in Scheme 5.

Step 1:

To a stirred solution of (E)-2-(3-iodostyryl)-1,3-dimethylbenzene (4)(0.551 g, 1.65 mmol) in NMP (10 mL) was added Et₃N (0.501 g, 4.95 mmol).The resulting solution was purged with nitrogen for 2 min andtris(dibenzylideneacetone)dipalladium(0) (0.076 g, 0.083 mmol) was addedfollowed by 1,1′-bis(diphenylphosphino)ferrocene (0.183 g, 0.330 mmol)After 0.5 h methyl thioglycolate (0.531 g, 5.00 mmol) was added and thereaction mixture was heated at 80° C. After 26 h the reaction mixturewas cooled to room temperature and diluted with water (50 mL). Theresulting mixture was extracted with ethyl acetate (3×50 mL) and thecombined organics were dried (Na₂SO₄), filtered and concentrated. Theresidue was purified by flash column chromatography (silica gel, 90:10hexanes/ethyl acetate) to provide(E)-2-(3-(2,6-dimethylstyryl)phenylthio)ethylamine (18) (0.473 g, 92%)as a colorless oil: ¹H NMR (500 MHz, CDCl₃) δ 7.54 (s, 1H), 7.34 (m,3H), 7.12 (d, J=16.4 Hz, 1H), 7.08 (m, 3H), 6.55 (d, J=16.4 Hz, 1H),3.73 (s, 3H), 3.69 (s, 2H), 2.36 (s, 6H).

Step 2:

To a stirred solution of 18 (0.259 g, 0.829 mmol) in THF (5 mL) at 0° C.was added lithium aluminum hydride (1.70 mL, 1M solution in diethylether, 1.70 mmol) at 0° C. After 10 min the reaction was quenched withsaturated aqueous ammonium chloride (0.25 mL), diluted with MTBE (100mL), the resulting suspension filtered and the filtrate concentrated.The resulting residue was purified by flash column chromatography(silica gel, 70:30 hexanes/ethyl acetate) to give 19 as a colorless oil.Yield (0.213 g, 90%): ¹H NMR (300 MHz, CDCl₃) δ 7.50 (s, 1H), 7.36-7.26(m, 3H), 7.11 (d, J=16.4 Hz, 1H), 7.07 (m, 3H), 6.54 (d, J=16.4 Hz, 1H),3.76 (q, J=6.0 Hz, 2H), 3.14 (t, J=6.0 Hz, 2H), 2.36 (s, 6H), 2.27 (t,J=6.0 Hz, 1H).

Step 3:

To a stirred solution of 19 (0.208 g, 0.731 mmol) and phthalimide (0.215g, 1.46 mmol) in THF (15 mL) at 0° C. was added diisopropylazodicarboxylate (0.365 g, 1.81 mmol), followed by solution oftriphenylphosphine (0.479 g, 1.83 mmol) in THF (2 mL). After 30 min thereaction mixture was diluted with brine (20 mL), extracted with hexanes(3×50 mL) and the combined extracts dried (Na₂SO₄), filtered andconcentrated. The resulting residue was purified by flash columnchromatography (silica gel, 90:10 hexanes/ethyl acetate) to give 20 as acolorless syrup. Yield (0.265 g, 88%): ¹H NMR (500 MHz, CDCl₃) δ 7.78(m, 2H), 7.67 (m, 2H), 7.55 (s, 1H), 7.32-7.19 (m, 3H), 7.17 (d, J=16.4Hz, 1H), 7.08 (m, 3H), 6.52 (d, J=16.4 Hz, 1H), 3.97 (t, J=6.8 Hz, 2H),3.27 (t, J=6.8 Hz, 2H), 2.39 (s, 6H).

Step 4:

A stirred solution of 20 (0.265 g, 0.641 mmol) and hydrazine monohydrate(0.096 g, 1.92 mmol) in methanol (10 mL) was heated at reflux for 2 h,cooled to room temperature and concentrated under reduced pressure. Theresidue was triturated with ethyl acetate (100 mL), the resultingsuspension filtered. The filtrate was concentrated under reducedpressure and the residue purified by flash column chromatography (silicagel, hexanes/ethyl acetate/7M ammonia in methanol (50:46:4) to provide(E)-2-(3-(2,6-dimethylstyryl)phenylthio)ethanamine as a colorless oil.Yield (0.163 g, 90%): R_(f) 0.58 (silica gel, 50:40:10 ethylacetate/hexanes/7N ammonia in methanol); ¹H NMR (500 MHz, CDCl₃) δ 7.52(s, 1H), 7.38 (m, 1H), 7.29 (m, 2H), 7.19 (d, J=16.4 Hz, 1H), 7.03 (s,3H), 6.56 (d, J=16.4 Hz, 1H), 3.05 (t, J=6.8 Hz, 2H), 2.82 (t, J=6.8 Hz,2H), 2.33 (s, 6H); ¹³C NMR (75 MHz, CDCl₃) δ 140.0, 138.1, 137.7, 137.3,134.7, 130.5, 129.9, 129.1, 129.0, 128.8, 128.0, 125.3, 41.7, 37.6,21.3; ESI MS m/z 284 [M+H]⁺; HPLC (Method 2) 98.5% (AUC), t_(R)=7.91min. HRMS calcd for C₁₈H₂₁NS [M+H−NH₃]: 267.1207. Found: 267.1195.

Example 19 Preparation of(E)-2-(3-(2,6-dimethylstyryl)phenylsulfinyl)ethanamine

(E)-2-(3-(2,6-dimethylstyryl)phenylsulfinyl)ethanamine was preparedfollowing method shown in scheme 6.

Step 1:

To a suspension of 20 (0.252 g, 0.610 mmol) in acetonitrile (1.5 mL) wasadded iron (III) chloride (0.003 g, 0.018 mmol) After 5 min periodicacid (0.168 g, 0.740 mmol) was added in one portion. The solution wasthen stirred for 10 min and 10% aqueous sodium thiosulfate (7 mL) wasadded. The resulting mixture was extracted with methylene chloride (3×20mL), the combined organics dried (MgSO₄), filtered and concentrated toprovide of 21 as a yellow oil. Yield (0.382 g, >99%): ¹H NMR (500 MHz,CDCl₃) δ 7.82-7.80 (m, 3H), 7.72-7.70 (m, 2H), 7.51-7.44 (m, 3H), 7.22(d, J=16.6 Hz, 1H), 7.11-7.08 (m, 3H), 6.59 (d, J=16.5 Hz, 1H),4.15-4.11 (m, 2H), 3.31-3.27 (m, 2H), 2.38 (s, 6H); ESI MS m/z 430[M+H]⁺.

Step 2:

Deprotection of Compound 21 following the method described in Example 18gave Example 19 as a yellow oil. Yield (0.115 g, 63%): R_(f) 0.56(silica gel, 90:10 Methylene Chloride/7N Ammonia in Methanol); ¹H NMR(500 MHz, CD₃OD) δ 7.87 (s, 1H), 7.75-7.73 (m, 1H), 7.61-7.58 (m, 2H),7.34 (d, J=16.7 Hz, 1H), 7.05 (s, 3H), 6.69 (d, J=16.7 Hz, 1H),3.12-3.08 (m, 2H), 3.04-2.96 (m, 2H), 2.35 (s, 6H); ¹³C NMR (125 MHz,CD₃OD) δ 143.2, 139.2, 136.1, 135.7, 132.3, 129.5, 129.0, 128.8, 127.5,126.6, 122.5, 121.0, 58.7, 35.1, 19.7; ESI MS m/z 300 [M+H]⁺; HPLC(Method 2) 97.4% (AUC), t_(R)=5.73 min. HRMS calcd for C₁₈H₂₁NOS [M+H]:300.1422. Found: 300.1409.

Example 20 Preparation of(E)-2-(3-(2,6-dimethylstyryl)phenylsulfonyl)ethanamine

(E)-2-(3-(2,6-dimethylstyryl)phenylsulfonyl)ethanamine was preparedfollowing method shown in scheme 7.

Step 1:

To a stirred solution of 20 (0.250 g, 0.60 mmol) at 0° C. was addedammonium molybdate tetrahydrate (0.226 g, 0.18 mmol) and hydrogenperoxide (0.50 mL, 35% solution, 5.70 mmol). After 30 min the solutionwas warmed to room temperature and stirred for an additional 16 h. Afterthis time the solution was cooled to 0° C. and quenched by addition of10% Na₂S₂O₃ (25 mL) and brine (25 mL). The resulting solution wasextracted with ethyl acetate (3×50 mL) and the combined organics weredried (MgSO₄), filtered and concentrated. Purification by flash columnchromatography (silica gel, 0-40% ethyl acetate/hexanes) provided 0.198g (74%) of 22 as a white solid: ¹H NMR (500 MHz, CDCl₃) δ 8.04 (s, 1H),7.81-7.78 (m, 3H), 7.70-7.68 (m, 2H), 7.59 (d, J=7.8 Hz, 1H), 7.48 (t,J=7.8 Hz, 1H), 7.26 (d, J=16.5 Hz, 1H), 7.12-7.09 (m, 3H), 6.60 (d,J=16.7 Hz, 1H), 4.11 (d, J=6.5 Hz, 2H), 3.65 (t, J=6.5 Hz, 2H), 2.40 (s,6H); ESI MS m/z 446 [M+H]⁺.

Step 2:

Deprotection of Compound 22 following the method described in Example 18gave Example 20 as a yellow semi-solid. Yield (0.113 g, 81%): R_(f) 0.70(silica gel, 90:10 Methylene Chloride/7N Ammonia in Methanol); ¹H NMR(500 MHz, CD₃OD) δ 8.05 (s, 1H), 7.93 (d, J=7.8 Hz, 1H), 7.82 (d, J=7.9Hz, 1H), 7.65 (t, J=7.8 Hz, 1H), 7.37 (d, J=16.7 Hz, 1H), 7.06 (s, 3H),6.72 (d, J=16.7 Hz, 1H), 3.39 (t, J=6.7 Hz, 2H), 2.98 (t, J=6.7 Hz, 2H),2.36 (s, 6H); ¹³C NMR (125 MHz, CD₃OD) δ 141.2, 140.7, 137.5, 137.2,133.3, 132.4, 131.2, 131.0, 129.0, 128.2, 127.7, 126.5, 58.9, 36.8,21.1; ESI MS m/z 316 [M+H]⁺; HPLC (Method 2) 98.4% (AUC), t_(R)=6.76min. HRMS calcd for C₁₈H₂₁NO₂S [M+H]: 316.1371. Found: 316.1372.

Example 21 Preparation of(E)-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine

(E)-3-(3-(2-(2,6,6-Trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-aminewas prepared by couplingtriphenyl((2,6,6-trimethylcyclohex-1-enyl)methyl)phosphonium bromide(24) (Scheme 8) with 3-(3-(1,3-dioxoisoindolin-2-yl)propyl)benzaldehyde(29) (Scheme 9).

Step 1:

To a stirred solution of 2,6,6-trimethylcyclohex-1-enecarbaldehyde (10.0g, 65.7 mmol) in anhydrous diethyl ether (100 mL) cooled to 0° C. wasadded lithium aluminum hydride (26.3 mL, 1M solution in THF, 26.3 mmol)dropwise over 10 min under nitrogen, the solution stirred at 0° C. for10 min and quenched with 2N aqueous sodium hydroxide (2 mL). Theresulting suspension diluted with MTBE (200 mL) and filtered. The filtercake was washed with MTBE (50 mL). The combined filtrate wasconcentrated under reduced pressure and the resulting residue dried invacuo to provide 23 as a white solid. Yield (9.28 g, 92%): ¹H NMR (500MHz, CDCl₃) δ 4.14 (d, J=3.4 Hz, 2H), 1.75 (s, 3H), 1.60 (m, 2H), 1.60(m, 2H), 1.44 (m, 2H), 1.04 (s, 6H), 0.97 (t, J=3.4 Hz, 1H); ESI MS m/z137 [M+H−H₂O]⁺.

Step 2:

To a stirred solution of triphenylphosphine hydrobromide (20.0 g, 58.3mmol) in MeOH (50 mL) was added a solution of 23 (8.99 g, 58.4 mmol) inmethanol (40 mL) and the reaction mixture stirred at room temperaturefor 18 h. The reaction solution was concentrated under reduced pressure,the residue triturated with a mixture of acetone (15 mL) and diethylether (200 mL) and the solvent decanted. The resulting residue was thentriturated with a mixture of acetone (10 mL), ethyl acetate (50 mL) anddiethyl ether (300 mL) and the solvent decanted again. The residue wasdried in vacuo to provide couplingtriphenyl((2,6,6-trimethylcyclohex-1-enyl)methyl)phosphonium bromide(26) as a white foam. Yield (21.9 g, 78%: ¹H NMR (500 MHz, DMSO-d₆) δ7.90-7.70 (m, 15H), 4.30 (d, J=15.1 Hz, 2H), 2.02 (m, 2H), 1.61 (d,J=5.7 Hz, 3H), 1.54 (m, 2H), 1.37 (m, 2H), 0.72 (s, 6H); ESI MS m/z 399[M−Br]⁺.

Step 3:

To a stirred solution of ethyl 3-(3-formylphenyl)propanoate (25) 6.78 g,32.9 mmol) in triethyl orthoformate (9.80 g, 66.1 mmol) was addedsulfamic acid (0.319 g, 3.29 mmol) at room temperature. After 67 h thereaction mixture was diluted with a mixture of MTBE and hexanes (1:1,100 mL), filtered and the filtrate concentrated. The residue waspurified by flash column chromatography (silica gel, 85:15 hexanes/ethylacetate) to give 26 as a colorless oil. Yield (7.31 g, 79%): ¹H NMR (500MHz, CDCl₃) δ 7.29 (m, 3H), 7.15 (d, J=7.3 Hz, 1H), 5.47 (s, 1H), 4.12(q, J=7.1 Hz, 2H), 3.63-3.50 (m, 4H), 2.96 (t, J=7.6 Hz, 2H), 2.62 (t,J=7.6 Hz, 2H), 1.23 (m, 9H)

Step 4:

Compound 26 was reduced following the procedure used in Example 18 togive 27 as a colorless oil. Yield (6.04 g, quant;): ¹H NMR (500 MHz,CDCl₃) δ 7.31-7.13 (m, 4H), 5.47 (s, 1H), 3.65-3.52 (m, 6H), 2.71 (t,J=7.6 Hz, 2H), 1.88 (m, 3H), 1.23 (m, 6H); ESI MS m/z 221 [M+H−H₂O]⁺.

Step 5:

Compound 27 was converted to the phthalimide following the procedureused in Example 18 to give 28 as a colorless oil. Yield (8.55 g, 92%):¹H NMR (300 MHz, CDCl₃) δ 7.83 (m, 2H), 7.70 (m, 2H), 7.29-7.14 (m, 4H),5.45 (s, 1H), 3.75 (t, J=7.2 Hz, 2H), 3.63-3.49 (m, 4H), 2.70 (t, J=7.6Hz, 2H), 2.03 (m, 2H), 1.23 (m, 6H).

Step 6:

To a stirred solution of 28 (8.55 g, 23.3 mmol) in 5:1 acetone/water (60mL) was added p-toluenesulfonic acid monohydrate (0.443 g, 2.33 mmol).After 4 h, the reaction mixture was concentrated under reduced pressure.The residue was dissolved in ethyl acetate (200 mL), the resultingsolution washed with saturated aqueous sodium bicarbonate (2×50 mL) andbrine (50 mL), dried (Na₂SO₄), filtered and concentrated under reducedpressure to provide 29 as an off-white solid (6.77 g, 99%): ¹H NMR (500MHz, CDCl₃) δ 9.97 (s, 1H), 7.82 (m, 2H), 7.71 (m, 3H), 7.66 (d, J=7.5Hz, 1H), 7.48 (d, J=7.6 Hz, 1H), 7.42 (t, J=7.5 Hz, 1H), 3.77 (t, J=7.1Hz, 2H), 2.78 (t, J=7.6 Hz, 2H), 2.08 (m, 2H); ESI MS m/z 294 [M+H]⁺.

Step 7:

Triphenyl((2,6,6-trimethylcyclohex-1-enyl)methyl)phosphonium bromide(24) was coupled with 3-(3-(1,3-dioxoisoindolin-2-yl)propyl)benzaldehyde(29) following the method described in Example 1 to give Compound 30 asa yellow oil (Scheme 11). Yield (0.283 g, 68%): ¹H NMR (500 MHz, CDCl₃)δ 7.82 (dd, J=5.4, 3.1 Hz, 2H), 7.69 (dd, J=5.5, 3.0 Hz, 2H), 7.21-7.17(m, 3H), 7.05 (d, J=6.8 Hz, 1H), 6.65 (d, J=16.3 Hz, 1H), 6.28 (d,J=16.3 Hz, 1H), 3.77 (t, J=7.2 Hz, 2H), 2.69 (t, J=7.8 Hz, 2H),2.08-2.02 (m, 4H), 1.75 (s, 3H), 1.65-1.61 (m, 2H), 1.50-1.48 (m, 2H),1.06 (s, 6H); ESI MS m/z 414 [M+H]⁺.

Step 8:

Deprotection following the method used in Example 18 gave Example 22 asa yellow oil. Yield (0.888 g, 87%): ¹H NMR (500 MHz, CD₃OD) δ 7.23-7.19(m, 3H), 7.06-7.04 (m, 1H), 6.67 (d, J=16.4 Hz, 1H), 6.29 (d, J=16.3 Hz,1H), 2.67-2.62 (m, 4H), 2.05 (t, J=6.1 Hz, 2H), 1.79 (q, J=7.5 Hz, 2H),1.74 (s, 3H), 1.69-1.64 (m, 2H), 1.52-1.50 (m, 2H), 1.06 (s, 6H); ESI MSm/z 284 [M+H]⁺.

Example 22 was further purified by conversion to the oxalate salt usingthe following procedure: To a stirred solution of(E)-3-(3-(2-(2,6,6-Trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine(0.426 g, 1.50 mmol) in ethanol (3 mL) was added oxalic acid (2.0 mL,20% solution in ethanol) at room temperature. After 30 min thesuspension was filtered and the resulting solid dried to provide(E)-3-(3-(2-(2,6,6-Trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amineoxalate salt as a white solid (0.433 g, 71%): ¹H NMR (500 MHz, DMSO-d₆)δ 7.31 (d, J=7.6 Hz, 2H), 7.26 (t, J=7.6 Hz, 1H), 7.07 (d, J=7.5 Hz,1H), 6.72 (d, J=16.3 Hz, 1H), 6.32 (d, J=16.4 Hz, 1H), 2.79 (t, J=7.6Hz, 2H), 2.64 (t, J=7.7 Hz, 2H), 2.02 (t, J=6.1 Hz, 2H), 1.86 (quint,J=7.7 Hz, 2H), 1.73 (s, 3H), 1.65-1.58 (m, 2H), 1.48-1.45 (m, 2H), 1.05(s, 6H); ¹³C NMR (75 MHz, CD₃OD) δ 164.2, 141.2, 137.6, 137.1, 132.4,129.0, 128.7, 127.1, 126.9, 125.9, 123.7, 33.9, 32.4, 31.8, 28.8, 21.4,18.7; ESI MS m/z 284 [M+H]⁺; HPLC (Method 2) 98.3% (AUC), t_(R)=9.99min. HRMS calcd for C₂₀H₂₉N [M+H]: 284.2378. Found: 284.2364; Anal.Calcd for C₂₀H₂₉N.1.35; C₂H₄O₄: C, 67.32; H, 7.89; N, 3.46. Found: C,67.31; H, 8.12; N, 3.47.

Example 22 Preparation of(E)-3-(3-(2-cyclohexylvinyl)phenyl)propan-1-amine

Step 1:

Coupling of aldehyde 29 with (cyclohexylmethy)ltriphenylphosphoniumbromide following the method used in Example 1 gave2-(3-(3-(2-Cyclohexylvinyl)phenyl)propyl)isoindoline-1,3-dione as ayellow oil. Yield (0.118 g, 56%) Isomer ratio trans:cis>9:1: ¹H NMR (500MHz, CDCl₃) δ 7.84-7.81 (m, 2H), 7.72-7.68 (m, 2H), 7.20 (t, J=7.5 Hz,1H), 7.16-7.00 (m, 3H), 6.25 (d, J=11.8 Hz, 1H), 5.46 (t, J=10.8 Hz,1H), 3.76 (t, J=7.2 Hz, 2H), 2.68 (t, J=7.9 Hz, 2H), 2.65-2.51 (m, 1H),2.07-2.01 (m, 2H), 1.80-1.63 (m, 4H), 1.40-1.11 (m, 6H); ESI MS m/z 374[M+H]⁺.

Step 2:

Deprotection following the method used in Example 18 gave(E)-3-(3-(2-Cyclohexylvinyl)phenyl)propan-1-amine (0.066 g, 85%).Purification by Preparative HPLC (Method 1) provided Example 22 as ayellow oil (0.037 g, 48%): R_(f) 0.39 (silica gel, 95:5 MethyleneChloride/7N Ammonia in Methanol); ¹H NMR (500 MHz, CD₃OD) δ 7.22 (t,J=7.7 Hz, 1H), 7.06 (d, J=8.8 Hz, 3H), 6.29 (d, J=11.7 Hz, 1H), 5.45 (d,J=11.6 Hz, 1H), 2.68-2.62 (m, 4H), 2.58-2.52 (m, 1H), 1.82-1.65 (m, 6H),1.33-1.14 (m, 6H); ¹³C NMR (75 MHz, CD₃OD) δ 143.1, 139.6, 136.3, 129.7,129.2, 128.3, 127.7, 127.1, 42.0, 38.4, 35.4, 34.4, 34.2, 27.1, 26.9;ESI MS m/z 244 [M+H]⁺ HPLC (Method 2) 98.2% (AUC), t_(R)=9.78 min. HRMScalcd for C₁₇H₂₅N [M+H]: 244.2065. Found: 244.2056.

Example 23 Preparation of (E)-3-(3-(pent-1-enyl)phenyl)propan-1-amine

Step 1:

Coupling of aldehyde 29 with butyltriphenylphosphonium bromide followingthe method used in Example 1 gave2-(3-(3-(pent-1-enyl)phenyl)propyl)isoindoline-1,3-dione as a yellowoil. Yield (0.198 g, 46%), isomer ratio 5:1: ¹H NMR (500 MHz, CDCl₃) δ7.84-7.81 (m, 2H), 7.72-7.68 (m, 2H), 7.22-7.01 (m, 4H), 6.37-6.16 (m,1H), 5.66-5.61 (m, 1H), 3.76 (t, J=7.2 Hz, 2H), 2.69-2.65 (m, 2H),2.31-2.15 (m, 2H), 2.04 (quint, J=7.5 Hz, 2H), 1.51-1.40 (m, 2H),0.97-0.92 (m, 3H).

Step 2:

Deprotection following the methods used in Example 18 gave3-(3-(pent-1-enyl)phenyl)propan-1-amine (0.106 g, 88%) as a 3:1 mixtureof trans- and cis-isomers. A portion of the sample was purified usingPreparative HPLC (Method 1) to provide Example 23 as a yellow oil: R_(f)0.75 (silica gel, 90:10 methylene chloride/7N ammonia in methanol); ¹HNMR (500 MHz, CD₃OD) δ 7.22 (t, J=7.6 Hz, 1H), 7.09-7.04 (m, 3H), 6.39(d, J=11.6 Hz, 1H), 5.64 (dt, J=11.7, 7.3 Hz, 1H), 2.67-2.62 (m, 4H),2.31-2.26 (m, 2H), 1.78 (quint, J=7.4 Hz, 2H), 1.47, (hex, J=7.4 Hz,2H), 0.93 (t, J=7.4 Hz, 3H); ¹³C NMR (125 MHz, CD₃OD) δ 143.1, 139.2,133.5, 130.3, 130.0, 129.2, 127.7, 127.3, 42.1, 35.7, 34.3, 21.8, 24.2,14.2; ESI MS m/z 204 [M+H]⁺; HPLC (Method 2)>99% (AUC), t_(R)=6.41 min.HRMS calcd for C₁₄H₂₁N [M+H]: 204.1752. Found: 204.1751.

Example 24 Preparation of (E)-3-(3-(hept-1-enyl)phenyl)propan-1-amine

Step 1:

Coupling of aldehyde 29 with hexyltriphenylphosphonium bromide followingthe method used in Example 1 gave2-(3-(3-(hept-1-enyl)phenyl)propyl)isoindoline-1,3-dione as a yellowoil. Yield (0.297 g, 68%), isomer ratio 5:1: ¹H NMR (500 MHz, CDCl₃) δ7.84-7.80 (m, 2H), 7.72-7.68 (m, 2H), 7.22-7.11 (m, 2H), 7.09-7.01 (m,2H), 6.35-6.18 (m, 1H), 5.67-5.61 (m, 1H), 3.75 (t, J=7.2 Hz, 2H), 2.67(q, J=7.6 Hz, 2H), 2.32-2.16 (m, 2H), 2.04 (quint, J=7.4 Hz, 2H),1.48-1.40 (m, 2H), 1.34-1.26 (m, 4H), 0.92-0.86 (m, 3H).

Step 2:

Deprotection following the methods used in Example 18 gave3-(3-(hent-1-enyl)phenyl)propan-1-amine (0.141 g, 74%) as a mixture oftrans- and cis-isomers. A portion of the sample was purified usingPreparative HPLC (Method 1) to provide Example 24 as a yellow oil: R_(f)0.71 (silica gel, 90:10 Methylene Chloride/7N Ammonia in Methanol); ¹HNMR (500 MHz, CD₃OD) δ 7.22 (t, J=7.6 Hz, 1H), 7.09-7.05 (m, 3H), 6.38(d, J=11.7 Hz, 1H), 5.66-5.61 (m, 1H), 2.64 (q, J=8.1 Hz, 4H), 2.32-2.27(m, 2H), 1.78 (quint, J=7.6 Hz, 2H), 1.45, (quint, J=7.4 Hz, 2H),1.34-1.30 (m, 4H), 0.90-0.87 (m, 3H); ¹³C NMR (125 MHz, CD₃OD) δ143.1,139.2, 133.7, 130.2, 129.9, 129.1, 127.6, 127.3, 42.1, 35.6, 34.2, 32.7,30.7, 29.6, 23.6, 14.4; ESI MS m/z 232 [M+H]⁺; HPLC (Method 2)>99%(AUC), t_(R)=8.79 min. HRMS calcd for C₁₆H₂₅N [M+H]: 232.2065. Found:232.2060.

Example 25 Preparation of(E)-3-amino-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenylpropan-1-ol

(E)-3-amino-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-olwas prepared following the method shown in Scheme 11.

Step 1:

Coupling of 3-iodobenzaldehyde with 24 following the method used inExample 1 gave 1-Iodo-3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)benzene(31) as a colorless oil. Yield (1.29 g, 85%): isomer ratio 9:1trans-/cis-isomers.

trans-isomer: ¹H NMR (500 MHz, CDCl₃) δ 7.74 (t, J=1.5 Hz, 1H), 7.53 (d,J=7.8 Hz, 1H), 7.34 (d, J=7.8 Hz, 1H), 7.04 (t, J=7.8 Hz, 1H), 6.66 (d,J=16.2 Hz, 1H), 6.23 (d, J=16.2 Hz, 1H), 2.03 (t, J=6.2 Hz, 2H), 1.74(s, 3H), 1.63 (m, 2H), 1.48 (m, 2H), 1.05 (s, 6H);

cis-isomer: ¹H NMR (500 MHz, CDCl₃) δ 7.88 (t, J=1.5 Hz, 1H), 7.47 (d,J=7.8 Hz, 1H), 7.34 (d, J=7.8 Hz, 1H), 6.98 (t, J=7.8 Hz, 1H), 6.28 (d,J=12.4 Hz, 1H), 6.13 (d, J=12.4 Hz, 1H), 1.96 (t, J=6.2 Hz, 2H), 1.66(m, 2H), 1.54 (m, 2H), 1.52 (s, 3H), 1.05 (s, 6H).

Step 2:

To a stirred solution of 31 (0.300 g, 0.852 mmol) in THF (10 mL) wasadded isopropylmagnesium chloride (0.46 mL, 2M solution in THF, 0.920mmol) at room temperature. After 1 h the reaction mixture was cooled to−20° C. and a solution of 3-(1,3-dioxoisoindolin-2-yl)propanal (32)(0.144 g, 0.709 mmol) in THF (3 mL) added. The resulting solution waswarmed to room temperature, quenched with brine (20 mL) and extractedwith ethyl acetate (3×50 mL). The combined organics were dried (Na₂SO₄),filtered and concentrated to give 33 which was used without purificationin the next step

Step 3:

Compound 33 was dissolved in methanol (15 mL) and hydrazine monohydrate(0.086 g, 1.71 mmol) added and the reaction mixture was heated atreflux. After 5 h the reaction mixture was cooled to room temperature,concentrated to an oil. The residue was purified by flash columnchromatography (silica gel, 50:40:10 ethyl acetate/hexanes/7N ammonia inmethanol) to provide Example 25 as a colorless oil. Yield (0.060 g,24%): R_(f) 0.17 (silica gel, 50:40:10 ethyl acetate/hexanes/7N ammoniain methanol); ¹H NMR (500 MHz, CD₃OD) δ 7.42 (s, 1H), 7.31-7.20 (m, 3H),6.72 (dd, J=16.2, 0.6 Hz, 1H), 6.32 (d, J=16.2 Hz, 1H), 4.73 (dd, J=8.1,5.2 Hz, 1H), 2.76 (m, 2H), 2.05 (t, J=6.2 Hz, 2H), 1.90 (m, 2H), 1.75(s, 3H), 1.66 (m, 2H), 1.52 (m, 2H), 1.07 (s, 6H); ¹³C NMR (125 MHz,CDCl₃) δ 146.9, 139.5, 139.1, 134.6, 130.5, 129.7, 128.8, 126.1, 125.9,124.5, 73.7, 42.8, 40.9, 39.8, 35.4, 34.0, 29.5, 22.0, 20.5; ESI MS m/z300 [M+H]⁺; HPLC (Method 2) 93.5% (AUC), t_(R)=9.72 min. HRMS calcd forC₂₀1-1₂₉NO [M+H]: 300.2327. Found: 300.2328.

Example 26 Preparation of(E)-2-methyl-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine

(E)-2-methyl-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-aminewas prepared following the method shown in Scheme 12.

Step 1:

To a stirred solution of 3-bromobenzaldehyde (2.00 g, 10.8 mmol), methylmethacrylate (1.35 g, 13.5 mmol), triethylamine (1.67 g, 16.5 mmol) andtriphenylphosphine (0.567 g, 2.16 mmol) in DMF (15 mL) was addedpalladium acetate (0.121 g, 0.539 mmol) and the reaction mixture washeated at 110° C. After 17 h the reaction mixture was cooled to roomtemperature, diluted with ethyl acetate (150 mL), the resultingsuspension washed with water (4×50 mL) and separated. The organic layerwas dried (Na₂SO₄), filtered and concentrated. The residue was purifiedby flash column chromatography (silica gel, 90:10 hexanes/ethyl acetate)to provide 34 as a light yellow oil. Yield (0.790 g, 36%): ¹H NMR (500MHz, CDCl₃) δ 10.05 (s, 1H), 7.89-7.47 (m, 4H), 5.54 (m, 1H), 3.84 (s,3H), 2.13 (s, 3H).

Step 2:

To a stirred solution of ester 34 (0.420 g, 2.06 mmol) and nickel(II)chloride hexahydrate (0.489 g, 2.06 mmol) in methanol (30 mL) was addedsodium borohydride (0.234 g, 6.19 mmol) portionwise over 5 min at roomtemperature. After the addition was complete, the reaction mixture wasstirred for 5 min and quenched with saturated aqueous ammonium chloride(10 mL). The resulting suspension was filtered through a short pad ofdiatomaceous earth and washed with methanol (50 mL). The combinedfiltrates were evaporated to dryness and the residue was partitionedbetween ethyl acetate (100 mL) and saturated aqueous ammonium chloride(50 mL). The organic layer was separated, dried (Na₂SO₄), filtered andconcentrated. The residue was dried in vacuo to provide 35 (0.417 g,97%) as a colorless oil: ¹H NMR (300 MHz, CDCl₃) δ 7.30-7.08 (m, 4H),4.67 (s, 2H), 3.64 (s, 3H), 3.04 (m, 1H), 2.81-2.63 (m, 2H), 1.15 (d,J=6.8 Hz, 3H).

Step 3:

To a stirred solution of oxalyl chloride (0.381 g, 3.00 mmol) inanhydrous methylene chloride (8 mL) cooled to −78° C. was addedanhydrous DMSO (0.703 g, 9.00 mmol) After 30 min a solution of 35 (0.417g, 2.00 mmol) in anhydrous methylene chloride (10 mL) was added, thesolution stirred for an additional 30 min, andN,N′-diisopropylethylamine (1.29 g, 10.0 mmol) was added. The resultingsolution was warmed to room temperature, quenched with saturated aqueousammonium chloride (20 mL), and extracted with methylene chloride (3×50mL). The combined organic layers were dried (Na₂SO₄), filtered andconcentrated under reduced pressure. The resulting residue was purifiedby flash column chromatography (silica gel, 90:10 hexanes/ethyl acetate)to provide 36 as a colorless oil. Yield (0.288 g, 70%): ¹H NMR (500 MHz,CDCl₃) δ 10.00 (s, 1H), 7.73 (m, 1H), 7.69 (s, 1H), 7.45 (m, 2H), 3.64(s, 3H), 3.09 (m, 1H), 2.77 (m, 2H), 1.18 (d, J=6.8 Hz, 3H).

Step 4:

Coupling of aldehyde 36 with Wittig reagent 24 following the method usedin Example 1 gave (E)-ethyl2-methyl-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propanoate(37) as a colorless oil. Yield (0.368 g, 81%): ¹H NMR (500 MHz, CDCl₃) δ7.26-7.16 (m, 3H), 7.01 (m, 1H), 6.65 (dd, J=16.3, 0.85 Hz, 1H), 6.30(d, J=16.3 Hz, 1H), 3.65 (s, 3H), 3.03 (m, 1H), 2.76 (m, 1H), 2.65 (m,1H), 2.03 (t, J=6.0 Hz, 2H), 1.75 (s, 3H), 1.64 (m, 2H), 1.49 (m, 2H),1.17 (d, J=6.9 Hz, 3H), 1.06 (s, 6H).

Step 5:

Reduction of ester 37 following the method used in Example 18 gavealcohol 38 as a colorless oil. Yield (0.327 g, 97%): ¹H NMR (500 MHz,CDCl₃) δ 7.24 (m, 2H), 7.19 (s, 1H), 7.03 (d, J=6.8 Hz, 1H), 6.67 (d,J=16.3 Hz, 1H), 6.32 (d, J=16.3 Hz, 1H), 3.58-3.48 (m, 2H), 2.75 (dd,J=13.5, 6.4 Hz, 1H), 2.42 (dd, J=13.5, 8.0 Hz, 1H), 2.04 (t, J=6.0 Hz,2H), 1.98 (m, 1H), 1.76 (s, 3H), 1.64 (m, 2H), 1.49 (m, 2H), 1.28 (t,J=5.7 Hz, 1H), 1.06 (s, 6H), 0.94 (d, J=6.9 Hz, 3H).

Step 6:

Alcohol 38 was converted to phthalimide 39 following the method used inExample 18 to give 39 as a colorless oil. Yield (0.419 g, 89%): ¹H NMR(500 MHz, CDCl₃) δ 7.78 (m, 2H), 7.67 (m, 2H), 7.15 (m, 3H), 7.00 (d,J=7.1 Hz, 1H), 6.63 (d, J=16.3 Hz, 1H), 6.26 (d, J=16.3 Hz, 1H), 3.67(dd, J=13.7, 7.0 Hz, 1H), 3.58 (dd, J=13.6, 7.2 Hz, 1H), 2.69 (m, 1H),2.47 (m, 2H), 2.04 (t, J=6.0 Hz, 2H), 1.76 (s, 3H), 1.64 (m, 2H), 1.49(m, 2H), 1.07 (s, 6H), 0.92 (d, J=6.9 Hz, 3H).

Step 7:

Deprotection of phthalimide 39 following the method used in Example 18gave(E)-3-(3-(2-(2,6,6-Trimethylcyclohex-1-enyl)vinyl)phenyl)butan-1-amineas a colorless oil. Yield (0.105 g, 36%): R_(f) 0.58 (silica gel,50:40:10 ethyl acetate/hexanes/7N ammonia in methanol); ¹H NMR (500 MHz,CD₃OD) δ 7.21 (m, 3H), 7.03 (dt, J=6.8, 1.6 Hz, 1H), 6.68 (dd, J=16.4,0.83 Hz, 1H), 6.30 (d, J=16.4 Hz, 1H), 2.70 (dd, J=13.4, 6.3 Hz, 1H),2.61 (dd, J=12.7, 5.7 Hz, 1H), 2.47 (dd, J=12.7, 7.1 Hz, 1H), 2.37 (dd,J=13.4, 8.2 Hz, 1H), 2.05 (t, J=6.1 Hz, 2H), 1.84 (m, 1H), 1.75 (s, 3H),1.66 (m, 2H), 1.51 (m, 2H), 1.06 (s, 6H), 0.90 (d, J=6.7 Hz, 3H); ¹³CNMR (125 MHz, CDCl₃) δ 142.6, 139.4, 139.1, 134.7, 130.4, 129.6, 129.1,128.5, 128.0, 124.7, 48.6, 42.1, 40.9, 39.2, 35.4, 34.0, 29.5, 22.0,20.5, 17.9; ESI MS m/z 298 [M+H]⁺; HPLC (Method 2) 98.4% (AUC),t_(R)=11.5 min. HRMS calcd for C₂₁H₃₁N [M+H]: 298.2535. Found: 298.2542.

Example 27 Preparation of(E)-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-1-amine

(E)-3-(3-(2-(2,6,6-Trimethylcyclohex-1-enyl)vinyl)phenyl)butan-1-aminewas prepared following the method used in Example 26.

Step 1:

Coupling of 3 bromobenzaldehyde with ethyl crotonate following themethod used in Example 26 gave ethyl 3-(3-formylphenyl)but-2-enoate as alight yellow oil Yield (0.547, 23%): ¹H NMR (300 MHz, CDCl₃) δ 10.05 (s,1H), 7.99 (t, J=1.6 Hz, 1H), 7.87 (dt, J=7.6, 1.3 Hz, 1H), 7.74 (m, 1H),7.56 (t, J=7.6 Hz, 1H), 6.19 (q, J=1.2 Hz, 1H), 4.23 (q, J=7.2 Hz, 2H),2.61 (d, J=1.2 Hz, 3H), 1.33 (t, J=7.2 Hz, 3H).

Step 2:

Reduction of ethyl 3-(3-formylphenyl)but-2-enoate following the methodused in Example 26 gave ethyl 3-(3-(hydroxymethyl)phenyl)butanoate as acolorless oil. Yield (0.427 g, 95%): ¹H NMR (300 MHz, CDCl₃) δ 7.32-7.15(m, 4H), 4.68 (s, 2H), 4.07 (q, J=7.2 Hz, 2H), 3.29 (m, 1H), 2.56 (m,2H), 1.30 (d, J=7.0 Hz, 3H), 1.19 (t, J=7.2 Hz, 3H).

Step 3:

Oxidation of ethyl 3-(3-(hydroxymethyl)phenyl)butanoate following themethod used in Example 26 gave ethyl 3-(3-formylphenyl)butanoate as acolorless oil. Yield (0.307 g, 68%): ¹H NMR (300 MHz, CDCl₃) δ 10.01 (s,1H), 7.73 (m, 2H), 7.50 (m, 2H), 4.06 (q, J=7.2 Hz, 2H), 3.37 (m, 1H),2.64 (dd, J=15.2, 7.4 Hz, 1H), 2.58 (dd, J=15.2, 7.6 Hz, 1H), 1.34 (d,J=7.0 Hz, 3H), 1.17 (t, J=7.2 Hz, 3H).

Step 4:

Coupling of ethyl 3-(3-formylphenyl)butanoate with Wittig reagent 24following the method used in Example 1 gave (E)-ethyl3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butanoate as acolorless oil. Yield (0.383 g, 81%): ¹H NMR (500 MHz, CDCl₃) δ 7.25 (m,3H), 7.07 (m, 1H), 6.66 (dd, J=16.3, 0.68 Hz, 1H), 6.31 (d, J=16.3 Hz,1H), 4.09 (q, J=7.0 Hz, 2H), 3.28 (m, 1H), 2.65-2.52 (m, 2H), 2.04 (t,J=6.2 Hz, 2H), 1.76 (s, 3H), 1.64 (m, 2H), 1.49 (m, 2H), 1.30 (d, J=6.9Hz, 3H), 1.20 (t, J=7.0 Hz, 3H), 1.06 (s, 6H).

Step 5:

Reduction of (E)-ethyl3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butanoate followingthe method used in Example 18 gave(E)-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-1-ol as acolorless oil. Yield (0.312 g, 93%): ¹H NMR (500 MHz, CDCl₃) δ 7.27 (m,2H), 7.20 (s, 1H), 7.03 (m, 1H), 6.67 (d, J=16.3 Hz, 1H), 6.32 (d,J=16.3 Hz, 1H), 3.60 (m, 2H), 2.89 (m, 1H), 2.04 (t, J=6.0 Hz, 2H), 1.88(q, J=7.3 Hz, 2H), 1.76 (s, 3H), 1.64 (m, 2H), 1.49 (m, 2H), 1.29 (d,J=7.0 Hz, 3H), 1.14 (t, J=5.3 Hz, 1H), 1.07 (s, 6H).

Step 6:

Conversion of(E)-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-1-ol withphthalimide following the method used in Example 18 gave(E)-2-(3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butyl)isoindoline-1,3-dioneas a colorless oil. Yield (0.416 g, 93%): ¹H NMR (500 MHz, CDCl₃) δ 7.75(m, 2H), 7.64 (m, 2H), 7.18 (s, 1H), 7.16 (t, J=7.6 Hz, 1H), 7.10 (d,J=7.7 Hz, 1H), 7.05 (d, J=7.5 Hz, 1H), 6.64 (d, J=16.3 Hz, 1H), 6.27 (d,J=16.3 Hz, 1H), 3.67 (t, J=7.1 Hz, 2H), 2.78 (m, 1H), 2.12 (m, 1H), 2.04(t, J=6.0 Hz, 2H), 1.93 (m, 1H), 1.77 (s, 3H), 1.64 (m, 2H), 1.49 (m,2H), 1.30 (d, J=7.0 Hz, 3H), 1.08 (s, 6H).

Step 7:

Deprotection of(E)-2-(3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butyl)isoindoline-1,3-dionefollowing the method used in Example 18 gave(E)-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-1-amineas a colorless oil. Yield (0.196 g, 68%), R_(f) 0.38 (silica gel,50:40:10 ethyl acetate/hexanes/7N ammonia in methanol); ¹H NMR (500 MHz,CDCl₃) δ 7.23 (m, 3H), 7.06 (m, 1H), 6.68 (dd, J=16.4, 0.83 Hz, 1H),6.30 (d, J=16.4 Hz, 1H), 2.77 (m, 1H), 2.58-2.48 (m, 2H), 2.05 (t, J=6.1Hz, 2H), 1.78 (m, 2H), 1.75 (s, 3H), 1.66 (m, 2H), 1.50 (m, 2H), 1.26(d, J=7.0 Hz, 3H), 1.07 (s, 6H); ¹³C NMR (75 MHz, CDCl₃) δ 148.8, 139.6,139.1, 134.8, 130.4, 129.9, 128.5, 126.8, 126.0, 124.8, 42.3, 41.1,40.9, 39.2, 35.5, 34.0, 29.5, 23.1, 22.0, 20.5; ESI MS m/z 298 [M+H]⁺;HPLC (Method 2) 97.5% (AUC), t_(R)=11.5 min. HRMS calcd for C₂₁H₃₁N[M+H]: 298.2535. Found: 298.2527.

Example 28 Preparation of(E)-3-(3-(2,6-dimethylstyryl)-2-methylphenyl)propan-1-amine

(E)-3-(3-(2,6-Dimethylstyryl)-2-methylphenyl)propan-1-amine was preparedfollowing the method shown in Scheme 13.

Step 1:

To a stirred solution of 3-iodo-2-methylbenzoic acid (40) (5.00 g, 19.1mmol) and trimethyl borate (7 mL) in THF (5 mL) was addedborane-dimethyl sulfide complex (11.5 mL, 2M solution in THF, 23.0 mmol)dropwise at such rate that after gas evolution stopped it remained at agentle reflux and after the addition was complete the reaction mixturewas cooled to room temperature. After 1.5 h the reaction mixture wasquenched by slow addition of methanol (10 mL), the resulting mixture wasconcentrated, and the residue was dissolved in methylene chloride (100mL). The resulting solution was washed with 2M aqueous sodium hydroxide(80 mL) and water (100 mL), and the organic layer was dried (Na₂SO₄),filtered and concentrated. The residue was purified by flash columnchromatography (silica gel, 60:36:4 methylene chloride/hexanes/MTBE) togive 41 as a white solid. Yield (4.36 g, 92%): ¹H NMR (500 MHz, CDCl₃) δ7.79 (d, J=7.9 Hz, 1H), 7.34 (d, J=7.9 Hz, 1H), 6.89 (t, J=7.9 Hz, 1H),4.72 (d, J=5.8 Hz, 2H), 2.47 (s, 3H), 1.58 (t, J=5.8 Hz, 1H).

Step 2:

Alcohol 41 was oxidized following the method used in Example 26 to givealdehyde 42 as a light yellow solid. Yield (1.39 g, 93%): ¹H NMR (300MHz, CDCl₃) δ 10.19 (s, 1H), 8.07 (d, J=7.9 Hz, 1H), 7.79 (d, J=7.9 Hz,1H), 7.09 (t, J=7.9 Hz, 1H), 2.79 (s, 3H).

Step 3:

Aldehyde 42 was coupled with Wittig reagent 3 following the method usedin Example 1 to give olefin 43 as a white semi-solid. Yield (0.641 g,65%), isomer ratio 4:1 trans:cis: ¹H NMR (300 MHz, CDCl₃) δ 7.80-6.45(m, 8H), 2.51-2.07 (m, 9H).

Step 4:

Coupling of olefin 43 with allyl alcohol following the method used inExample 1 gave aldehyde 44 as a light yellow oil. Yield (0.398 g, 86%),isomer ratio 4:1 trans:cis: ¹H NMR (500 MHz, CDCl₃) δ 9.83 (m, 1H),7.49-6.58 (m, 8H), 2.98 (m, 2H), 2.72 (m, 2H), 2.45-2.07 (m, 9H); ESI MSm/z 261 [M+H−H₂O]⁺.

Step 5:

Reductive amination of aldehyde 44 with ammonia following the methodused in Example 1 followed by purification by Preparative HPLC(Method 1) gave(E)-3-(3-(2,6-Dimethylstyryl)-2-methylphenyl)propan-1-amine as a lightyellow oil. Yield (0.038 g, 10%): R_(f) 0.55 (silica gel, 50:40:10 ethylacetate/hexanes/7N ammonia in methanol); ¹H NMR (500 MHz, CD₃OD) δ 7.43(dd, J=7.5, 1.3 Hz, 1H), 7.10 (m, 2H), 7.03 (s, 3H), 6.91 (d, J=16.6 Hz,1H), 6.87 (d, J=16.6 Hz, 1H), 2.70 (t, J=7.4 Hz, 4H), 2.37 (s, 6H), 2.32(s, 3H), 1.74 (m, 2H); ¹³C NMR (125 MHz, CD₃OD) δ 141.9, 139.3, 138.7,137.1, 134.7, 134.6, 130.0, 129.9, 129.0, 127.8, 127.0, 125.3, 42.5,34.8, 32.5, 21.4, 15.5; ESI MS m/z 280 [M+H]⁺; HPLC (Method 5)>99%(AUC), t_(R)=12.11 min. HRMS Calcd for C₂₀H₂₅N [M+H]: 280.2065. Found:280.2052.

Example 29 Preparation of(Z)-3-(3-(2,6-dimethylstyryl)-2-methylphenyl)propan-1-amine

(V)-3-(3(2,6-Dimethylstyryl)-2-methylphenyl)propan-1-amine as wasisolated during the synthesis of Example 28 as a light yellow oil. Yield(0.036 g, 9%): R_(f) 0.55 (silica gel, 50:40:10 ethyl acetate/hexanes/7Nammonia in methanol); ¹H NMR (500 MHz, CD₃OD) δ 6.98 (t, J=7.8 Hz, 1H),6.93 (d, J=7.5 Hz, 1H), 6.90 (d, J=7.5 Hz, 2H), 6.87 (d, J=12.1 Hz, 1H),6.68 (t, J=7.6 Hz, 1H), 6.61 (d, J=12.1 Hz, 1H), 6.49 (d, J=7.6 Hz, 1H),2.67 (t, J=7.4 Hz, 2H), 2.66 (t, J=7.3 Hz, 2H), 2.30 (s, 3H), 2.05 (s,6H), 1.70 (m, 2H); ¹³C NMR (125 MHz, CD₃OD) δ 141.6, 138.7, 137.9,136.9, 134.6, 132.2, 129.9, 129.4, 128.5, 127.8, 127.5, 126.1, 42.5,34.7, 32.4, 20.6, 15.6; ESI MS m/z 280 [M+H]⁺; HPLC (Method 5)>99%(AUC), t_(R)=12.43 min. HRMS Calcd for C₂₀H₂₅N [M+H]: 280.2065. Found:280.2053.

Example 30 Preparation of(E)-2-amino-N-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)acetamide

(E)-2-amino-N-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)acetamidewas prepared following the method used in Example 15.

Step 1:

Coupling of Wittig reagent 24 with 3-nitrobenzaldehyde gave1-nitro-3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)benzene as a lightyellow oil. Yield (0.639 g, 95%), isomer ratio 4:1 ratio trans:cis.

trans-isomer: ¹H NMR (300 MHz, CDCl₃) δ 8.24 (t, J=1.9 Hz, 1H), 8.04 (m,1H), 7.69 (d, J=7.7 Hz, 1H), 7.47 (t, J=8.0 Hz, 1H), 6.83 (dd, J=16.3,0.85 Hz, 1H), 6.40 (d, J=16.3 Hz, 1H), 2.06 (t, J=6.2 Hz, 2H), 1.81 (s,3H), 1.65 (m, 2H), 1.52 (m, 2H), 1.08 (s, 6H);

cis-isomer: ¹H NMR (300 MHz, CDCl₃) δ 8.48 (t, J=1.9 Hz, 1H), 8.00 (m,1H), 7.65 (d, J=7.7 Hz, 1H), 7.41 (t, J=8.0 Hz, 1H), 6.46 (d, J=12.4 Hz,1H), 6.29 (d, J=12.4 Hz, 1H), 1.98 (m, 2H), 1.65 (m, 2H), 1.57 (m, 2H),1.41 (s, 3H), 1.08 (s, 6H).

Step 2:

Reduction of 1-nitro-3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)benzenefollowing the method described in Example 15 gave3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)aniline as a light yellowoil. Yield (0.639 g, 95%), isomer ratio 4:1 trans:cis. trans-isomer: ¹HNMR (300 MHz, CDCl₃) δ 7.11 (t, J=7.8 Hz, 1H), 6.82 (d, J=7.7 Hz, 1H),6.75 (t, J=2.0 Hz, 1H), 6.63 (dd, J=16.3, 0.85 Hz, 1H), 6.56 (ddd,J=7.9, 2.3, 0.80 Hz, 1H), 6.25 (d, J=16.3 Hz, 1H), 3.65 (br s, 2H), 2.03(t, J=6.2 Hz, 2H), 1.75 (s, 3H), 1.63 (m, 2H), 1.50 (m, 2H), 1.05 (s,6H);

cis-isomer: ¹H NMR (300 MHz, CDCl₃) δ 7.04 (t, J=7.8 Hz, 1H), 6.88 (d,J=7.7 Hz, 1H), 6.77 (t, J=2.0 Hz, 1H), 6.53 (m, 1H), 6.30 (d, J=12.4 Hz,1H), 6.04 (d, J=12.4 Hz, 1H), 3.54 (br s, 2H), 1.96 (m, 2H), 1.63 (m,2H), 1.53 (m, 2H), 1.44 (s, 3H), 1.07 (s, 6H).

Step 3:

Amidation of 3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)anilinefollowing the method described in Example 15 gave(9H-Fluoren-9-yl)methyl2-oxo-2-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenylamino)ethylcarbamateas a white foam. Yield (0.175 g, 74%), isomer ratio 10:1 trans: cis;

trans-isomer: ¹H NMR (500 MHz, CDCl₃) δ 7.76 (m, 3H), 7.59 (m, 2H), 7.51(s, 1H), 7.40 (m, 3H), 7.30 (m, 3H), 7.18 (m, 1H), 6.69 (d, J=16.3 Hz,1H), 6.31 (d, J=16.3 Hz, 1H), 5.46 (br s, 1H), 4.50 (m, 2H), 4.24 (t,J=6.7 Hz, 1H), 4.00 (br s, 2H), 2.04 (t, J=6.2 Hz, 2H), 1.75 (s, 3H),1.64 (m, 2H), 1.48 (m, 2H), 1.06 (s, 6H);

cis-isomer: ¹H NMR (500 MHz, CDCl₃) δ 7.76 (m, 3H), 7.59 (m, 2H), 7.51(s, 1H), 7.40 (m, 3H), 7.30 (m, 3H), 7.18 (m, 1H), 6.36 (d, J=12.4 Hz,1H), 6.12 (d, J=12.4 Hz, 1H), 5.46 (br s, 1H), 4.50 (m, 2H), 4.24 (t,J=6.7 Hz, 1H), 4.00 (br s, 2H), 1.96 (m, 2H), 1.64 (m, 2H), 1.52 (m,2H), 1.42 (s, 3H), 1.06 (s, 6H).

Step 4:

Deprotection of (9H-Fluoren-9-yl)methyl2-oxo-2-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenylamino)ethylcarbamatefollowing the method used in Example 15, followed by purification byPreparative HPLC (Method 1) gave(E)-2-amino-N-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)acetamideas a colorless semi-solid. Yield (0.039 g, 39%): R_(f) 0.53 (silica gel,50:40:10 ethyl acetate/hexanes/7N ammonia in methanol); ¹H NMR (500 MHz,CDCl₃) δ 7.68 (s, 1H), 7.41 (d, J=8.0 Hz, 1H), 7.26 (t, J=7.8 Hz, 1H),7.14 (d, J=7.7 Hz, 1H), 6.71 (d, J=16.3 Hz, 1H), 6.30 (d, J=16.3 Hz,1H), 3.42 (s, 2H), 2.06 (t, J=6.2 Hz, 2H), 1.75 (s, 3H), 1.67 (m, 2H),1.51 (m, 2H), 1.07 (s, 6H); ¹³C NMR (125 MHz, CDCl₃) δ 173.9, 140.3,140.0, 139.0, 134.2, 130.7, 130.2, 129.3, 123.2, 119.9, 118.4, 45.8,40.9, 35.4, 34.0, 29.5, 22.0, 20.5; ESI MS m/z 299 [M+H]⁺; HPLC (Method2) 97.5% (AUC), t_(R)=9.99 min. HRMS calcd for C₁₉H₂₆N₂O [M+H]:299.2123. Found: 299.2125.

Example 31 Preparation of(E)-2-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenoxy)ethanamine

(E)-2-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenoxy)ethanamine wasprepared following the method shown in Scheme 14.

Step 1:

Coupling of 3-hydroxybenzaldehyde with Wittig reagent 24 following themethod used in Example 1 gave olefin 45 as a light yellow oil. Yield(0.543 g, 68%), isomer ratio 8:1 ratio trans:cis: ¹H NMR (300 MHz,CDCl₃) δ 7.19 (t, J=7.8 Hz, 1H), 6.97 (d, J=7.7 Hz, 1H), 6.89 (t, J=2.3Hz, 1H), 6.69 (m, 1H), 6.64 (dd, J=16.3, 0.87 Hz, 1H), 6.27 (d, J=16.3Hz, 1H), 2.03 (t, J=6.2 Hz, 2H), 1.75 (s, 3H), 1.62 (m, 2H), 1.48 (m,2H), 1.05 (s, 6H); The cis-isomer: ¹H NMR (300 MHz, CDCl₃) δ 7.12 (t,J=7.8 Hz, 1H), 7.01 (d, J=7.7 Hz, 1H), 6.92 (t, J=2.3 Hz, 1H), 6.69 (m,1H), 6.34 (d, J=12.4 Hz, 1H), 6.10 (d, J=16.3 Hz, 1H), 1.96 (m, 2H),1.62 (m, 2H), 1.48 (m, 2H), 1.43 (s, 3H), 1.07 (s, 6H).

Step 2:

To a stirred solution of olefin 45 (0.507 g, 2.09 mmol) and tert-butyl2-hydroxyethylcarbamate (1.35 g, 8.37 mmol) in THF (5 mL) was addedtriphenylphosphine (2.19 g, 8.35 mmol) followed by a solution ofdiisopropyl azodicarboxylate (1.69 g, 8.36 mmol) in THF (3 mL) dropwiseover 10 min at room temperature. After 70 h the reaction mixture washeated at 50 C for an additional 18 h. After this time the reactionmixture was cooled to room temperature, partitioned between brine (50mL) and ethyl acetate (100 mL). The organic layer was separated, dried(Na₂SO₄), filtered and concentrated. The resulting residue was purifiedby flash column chromatography (silica gel, 80:20 hexanes/ethyl acetate)to give 46 as a colorless syrup (0.731 g, 91%): ESI MS m/z 330[M+H−C₄H₈]⁺.

Step 3:

Deprotection of 46 following the method used in Example 13 gave(E)-2-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenoxy)ethanamine asa colorless oil. Yield (0.060 g, 11%): R_(f) 0.37 (silica gel, 50:40:10ethyl acetate/hexanes/7N ammonia in methanol); ¹H NMR (500 MHz, CDCl₃) δ7.21 (t, J=7.9 Hz, 1H), 6.99 (m, 2H), 6.81 (dd, J=7.8, 2.0 Hz, 1H), 6.69(dd, J=16.3, 0.79 Hz, 1H), 6.29 (d, J=16.3 Hz, 1H), 4.03 (t, J=5.3 Hz,2H), 3.01 (t, J=5.3 Hz, 2H), 2.05 (t, J=6.2 Hz, 2H), 1.75 (s, 3H), 1.66(m, 2H), 1.51 (m, 2H), 1.06 (s, 6H); ¹³C NMR (125 MHz, CDCl₃) δ 160.8,141.0, 139.0, 134.5, 130.7, 130.6, 129.0, 120.0, 114.5, 113.2, 70.4,42.1, 40.9, 35.4, 34.0, 29.5, 22.0, 20.5; ESI MS m/z 286 [M+H]⁺; HPLC(Method 2) 94.8% (AUC), t_(R)=10.2 min. HRMS calcd for C₁₉H₂₇NO [M+H]:286.2171. Found: 286.2162.

Example 32 Preparation of(E/Z)-3-(3-(2-ethyl-6-methylstyryl)phenyl)propan-1-amine

(E/Z)-3-(3-(2-Ethyl-6-methylstyryl)phenyl)propan-1-amine was preparedfollowing the method described in Example 21, except that phthalimide 29was synthesized according to Scheme 15.

Step 1:

To a solution of 3-iodobenzaldehyde (8.92 g, 38.5 mmol) in anhydrousMeOH (40 mL) was added trimethyl orthoformate (7 mL, 64 mmol) andp-toluenesulfonic acid hydrate (0.36 g, 1.9 mmol). The mixture wasstirred for 15 min, then partitioned between EtOAc and saturated aqueousNaHCO₃. The combined organics were washed with brine, dried over MgSO₄and concentrated under reduced pressure to give iodide 47 as a whitesolid. Yield (10.7 g, 100%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.71-7.73 (m,2H), 7.40 (d, J=8.0 Hz, 1H), 7.19-7.23 (m, 1H), 5.36 (s, 1H), 3.24 (s,6H).

Step 2:

A mixture of NaHCO₃ (11.63 g, 138.4 mmol) and tetrabutylammonium bromide(13.1 g, 40.64 mmol) in degassed DMF (˜95 mL, degassed by bubbling withargon for 20 min) was further degassed with argon for 5 min. Allylalcohol (4.71 g, 81.1 mmol) and iodide 47 (10.29 g, 37.0 mmol) wereadded and the mixture purged twice (alternately put under vacuum andargon). Pd(OAc)₂ (0.5116 g, 2.28 mmol) was added and the mixture purgedagain. The reaction mixture was heated at 60° C. under argon for 5.5 hthen cooled to room temperature. The mixture was concentrated underreduced pressure and the residue suspended in EtOAc and sonicated. Thesolids were removed by filtration and the filtrate partiallyconcentrated under reduced pressure. The residue was washed withsaturated aqueous NaHCO₃ and brine, treated with activated charcoal andMgSO₄ and concentrated under reduced pressure. Purification by flashchromatography (20% EtOAc-hexanes) gave aldehyde 48 as an oil. Yield(6.20 g, 80%): ¹H NMR (400 MHz, DMSO-d₆) δ 9.71 (s, 1H), 7.29 (t, J=8.0Hz, 1H), 7.19-7.24 (m, 3H), 5.34 (s, 1H), 3.23 (s, 6H), 2.88 (t, J=8.0Hz, 2H), 2.75-2.79 (m, 2H).

Step 3:

To a solution of aldehyde 48 (6.20 g, 29.8 mmol) in EtOH (absolute, 50mL) was added NaBH₄ (0.699 g, 18.5 mmol). The mixture was stirred for 15min then partially concentrated under reduced pressure. The residue waspartitioned between EtOAc and saturated aqueous NaHCO₃ and the combinedorganics were washed with water and brine. The solution was dried overMgSO₄ and concentrated under reduced pressure to give alcohol 49 as acolorless liquid. Yield (6.11 g, 98%).

Step 4:

To an ice cold solution of alcohol 49 (6.11 g, 29.06 mmol) in THF (100mL) was added phthalimide (4.53 g, 30.8 mmol), and triphenylphosphine(9.606 g, 36.6 mmol) Diethyl azodicarboxylate (6.702 g, 38.5 mmol) wasadded and the mixture stirred for 10 min. After warming to roomtemperature, the reaction mixture was concentrated under reducedpressure. 10% EtOAc-hexanes was added and the mixture sonicated. Thesolids were removed by filtration, washed with 20% EtOAc-heptane, andthe filtrate was concentrated under reduced pressure. Purification byflash chromatography (25-30% EtOAc-heptane) gave phthalimide 50 as awhite waxy solid. Yield (9.77 g, 99%): ¹H NMR (400 MHz, DMSO-d₆) δ7.80-7.89 (m, 4H), 7.15-7.28 (m, 4H), 5.32 (s, 1H), 3.60 (t, J=8.0 Hz,2H), 3.22 (s, 6H), 2.64 (t, J=8.0 Hz, 2H), 1.87-1.99 (m, 2H).

Step 5:

To a solution of phthalimide 50 (5.66 g, 16.67 mmol) in acetone-water(5:1, 50 mL) was added p-toluenesulfonic acid hydrate (0.2116 g, 1.11mmol). The mixture was stirred for 3 h then concentrated under reducedpressure. The residue was partitioned between EtOAc and water and thecombined organics were washed with brine, dried over MgSO₄ andconcentrated under reduced pressure to give phthalimide 29 as a whitesolid. Yield (4.48 g, 92%). The NMR data was consistent with datareported above.

Step 6:

Preparation of 2-ethyl-6-methylbenzylphosphonium bromide: To a solutionof 2-ethyl-6-methylbenzylbromide (0.432 g, 2.03 mmol) in toluene (10 mL)was added triphenylphosphine (0.6018 g, 2.29 mmol) The mixture washeated at 100° C. for 4 h. After cooling to room temperature, theprecipitate was collected by filtration and washed with toluene to give2-ethyl-6-methylbenzylphosphonium bromide as a solid. Yield (0.958 g,99%). ¹H NMR (400 MHz, DMSO-d₆) δ 7.90-7.92 (m, 3H), 7.69 (ddd, J=8.4,8.4, 3.6 Hz, 6H), 7.49-7.55 (m, 6H), 7.20 (ddd, J=7.6, 7.6, 2.8 Hz, 1H),7.02 (d, J=8.0 Hz, 1H), 6.94 (d, J=7.6 Hz, 1H), 4.91 (d, J=14.7 Hz, 2H),2.05 (q, J=6.7 Hz, 2H), 1.75 (s, 3H), 0.78 (t, J=6.7 Hz, 3H).

To an ice cold solution of 2-ethyl-6-methylbenzylphosphonium bromide(0.9473 g, 1.99 mmol) and 18-crown-6 (0.0769 g, 0.29 mmol) in CH₂Cl₂ (10mL) under argon was added potassium tert-butoxide (0.242 g, 2.16 mmol)The mixture was stirred at 0° C. for 2 h. An ice cold solution ofphthalimide 29 (0.394 g, 0.34 mmol) in CH₂Cl₂ (10 mL) was added and themixture was stirred at 0° C. for 45 min, then allowed to warm to roomtemperature and stirred for 1.5 h. The mixture was concentrated underreduced pressure then triturated with ˜10% EtOAc-heptane. Solids wereremoved by filtration and the filtrate concentrated under reducedpressure. Purification by flash chromatography (20 to 50% EtOAc-hexanesgradient) gave phthalimide 51 as an oil. Yield (0.505 g, 92%): ¹H NMR(400 MHz, DMSO-d₆) δ 7.78-7.86 (m, 4H), 7.34 (s, 1H), 7.34 (d, J=7.6 Hz,1H), 7.23 (t, J=8.0 Hz, 1H), 7.21 (d, J=16.8 Hz, 1H), 7.04-7.13 (m, 4H),6.54 (d, J=16.8 Hz, 1H), 3.62 (t, J=6.8 Hz, 2H), 2.64 (t, J=7.4 Hz, 4H),2.29 (s, 3H), 1.90-1.99 (m, 2H), 1.11 (t, J=6.8 Hz, 3H).

Step 7:

To a solution of phthalimide 51 (0.5049 g, 1.24 mmol) in EtOH (absolute,10 mL) was added hydrazine hydrate (0.2 mL, 4.1 mmol). The mixture wasstirred at room temperature for 1 h then heated to reflux for 2 h. Aftercooling to room temperature, the mixture was concentrated under reducedpressure. The residue was suspended in heptane and the solids removed byfiltration. The filtrate was concentrated under reduced pressure to giveExample 32 as a colorless oil. Yield (0.230 g, 66%), trans-/cis-isomerratio 5:1. Trans-isomer: ¹H NMR (400 MHz, DMSO-d₆) δ 7.38-7.39 (m, 2H),7.26 (t, J=7.6 Hz, 1H), 7.21 (d, J=16.8 Hz, 1H), 7.03-7.12 (m, 4H), 6.56(d, J=16.4 Hz, 1H), 2.60-2.68 (m, 4H), 2.54 (t, J=6.8 Hz, 2H), 2.29 (s,3H), 1.60-1.68 (m, 2H), 1.11 (t, J=6.8 Hz, 3H).

Example 33 Preparation of(E/Z)-3-(3-(2,5-dimethylstyryl)phenyl)propan-1-amine

(E/Z)-3-(3-(2,5-Dimethylstyryl)phenyl)propan-1-amine was preparedfollowing the method described in Example 32.

Step 1:

To a suspension of the crude2,5-dimethylmethylbenzyltriphenylphosphonium bromide in THF (10 mL) andCH₂Cl₂ (5 mL) were added potassium tert-butoxide (0.163 g, 1.45 mmol)and 18-crown-6 (0.163 g, 1.45 mmol) The mixture was stirred at roomtemperature under argon for 30 min then sonicated for 1 min. A solutionof phthalimide 29 (0.193 g, 0.658 mmol) in CH₂Cl₂ (2 mL) was added andthe mixture stirred at room temperature for 2 h. The mixture wasconcentrated under reduced pressure and the residue partitioned betweenEtOAc and saturated aqueous NH₄Cl. The combined organics were washedwith brine, dried over MgSO₄ and concentrated under reduced pressure.Purification by flash chromatography (20 to 50% EtOAc-hexanes) gave(E)-2-(3-(3-(2,5-dimethylstyryl)phenyl)propyl)isoindoline-1,3-dione asan oil. Yield (0.225 g, 86%), trans-/cis-isomer 1:1.5. Cis-isomer: ¹HNMR (400 MHz, CDCl₃) δ 7.69-7.76 (m, 4H), 6.91-7.44 (m, 7H) 6.63 (d,J=12.0 Hz, 1H), 6.57 (d, J=12.0 Hz, 1H), 3.81 (t, J=7.2 Hz, 2H), 2.75(t, J=7.6 Hz, 2H), 2.44 (s, 3H), 2.39 (s, 3H), 2.08-2.17 (m, 2H).

Step 2:

(E)-2-(3-(3-(2,5-dimethylstyryl)phenyl)propyl)isoindoline-1,3-dione wasdeprotected following the method used in Example 32 to afford Example 33as an oil. Trans-/cis-isomer 2:1. Trans-isomer: ¹H NMR (400 MHz, CD₃OD)δ 7.35-7.43 (m, 3H), 7.28 (t, J=8.0 Hz, 1H), 6.88-7.19 (m, 5H), 2.70 (t,J=8.0 Hz, 4H), 2.38 (s, 3H), 2.34 (s, 3H), 1.80-1.87 (m, 2H).

Example 34 Preparation of(E/Z)-3-(3-(2,4-dimethylstyryl)phenyl)propan-1-amine

(E/Z)-3-(3-(2,4-Dimethylstyryl)phenyl)propan-1-amine was preparedfollowing the method described in Example 32.

Step 1:

Phthalimide 29 was coupled with2,4-dimethylmethylbenzyltriphenylphosphonium bromide and purified byflash chromatography (20 to 50% EtOAc-hexanes gradient) to give(E)-2-(3-(3-(2,4-dimethylstyryl)phenyl)propyl)isoindoline-1,3-dione asan oil. Yield (0.3974 g, 92%), trans-/cis-isomer ratio 1.2:1.Cis-isomer: ¹H NMR (400 MHz, CDCl₃) δ 7.80-7.88 (m, 4H), 6.80-7.55 (m,7H), 6.62 (d, J=12.0 Hz, 1H), 6.57 (d, J=12.0 Hz, 1H), 3.63 (t, J=7.2Hz, 2H), 2.65 (t, J=7.2 Hz, 2H), 2.37 (s, 3H), 2.27 (s, 3H), 1.95(quint, J=7.2 Hz, 2H).

Step 2:

(E)-2-(3-(3-(2,4-dimethylstyryl)phenyl)propyl)isoindoline-1,3-dione wasdeprotected following the method used in Example 32 to afford Example 34as an oil. Yield (0.0422 g, 16%), trans-/cis-isomer ratio 1:2.cis-isomer: ¹H NMR (400 MHz, DMSO-d₆) δ 6.93-7.11 (m, 5H), 6.88 (t,J=6.4, 2H), 6.63 (d, J=12.0 Hz, 1H), 6.59 (d, J=12.0 Hz, 1H), 2.43 (t,J=6.8 Hz, 4H), 2.25 (s, 3H), 2.17 (s, 3H), 1.47 (quint, J=6.8 Hz, 2H),1.31 (br s, 2H).

Example 35 Preparation of(E)-3-(3-(2,4,6-trimethylstyryl)phenyl)propan-1-amine

(E)-3-(3-(2,4,6-Trimethylstyryl)phenyl)propan-1-amine was preparedaccording to the method used in Example 32.

Step 1:

Phthalimide 29 was coupled with2,4,6-trimethylbenzyltriphenylphosphonium bromide to give(E)-2-(3-(3-(2,4,6-trimethylstyryl)phenyl)propyl)isoindoline-1,3-dione.Yield (0.2485 g, 46%), trans-/cis-isomer ratio 4:1. Trans-isomer: ¹H NMR(400 MHz, CDCl₃) δ 7.84-7.87 (m, 2H), 7.72-7.74 (m, 2H), 7.26-7.36 (m,3H), 7.16 (s, 1H), 7.12 (d, J=16.4 Hz, 1H), 6.94 (s, 2H), 6.57 (d,J=16.8 Hz, 1H), 3.82 (t, J=7.2 Hz, 2H), 2.76 (t, J=7.2 Hz, 2H), 2.39 (s,6H), 2.33 (s, 3H), 2.10-2.17 (m, 2H).

Step 2:

(E)-2-(3-(3-(2,4,6-Trimethylstyryl)phenyl)propyl)isoindoline-1,3-dionewas deprotected to afford Example 35 as an oil. Yield (60%): ¹H NMR (400MHz, CD₃OD) δ 7.33-7.35 (m, 2H), 7.27 (t, J=7.2 Hz, 1H), 7.14 (d, J=16.4Hz, 1H), 7.12 (d, J=7.2 Hz, 1H), 6.87 (s, 2H), 6.55 (d, J=16.8 Hz, 1H),2.66-2.70 (m, 4H), 2.32 (s, 6H), 2.26 (s, 3H), 1.82 (quint, J=7.6 Hz,2H).

Example 36 Preparation of(E/Z)-3-(3-(2-ethylstyryl)phenyl)propan-1-amine

(E)-3-(3-(2-Ethylstyryl)phenyl)propan-1-amine was prepared according tothe method used in Example 32.

Step 1:

Phthalimide 29 was coupled with 2-ethylbenzyltriphenylphosphoniumbromide according to the method used in Example 32, except that thereaction was stirred at room temperature for 3.5 h. Purification byflash chromatography (20 to 50% EtOAc-hexanes gradient) gave(E/Z)-2-(3-(3-(2-ethylstyryl)phenyl)propyl)isoindoline-1,3-dione as anoil. Yield (0.4158 g, 83%), trans-/cis-isomer ratio 1:1. Cis-isomer: ¹HNMR (400 MHz, DMSO-d₆) δ 7.80-7.91 (m, 4H), 6.83-7.47 (m, 8H), 6.73 (d,J=12.0 Hz, 1H), 6.60 (d, J=12.0 Hz, 1H), 3.49 (t, J=7.2 Hz, 2H), 2.79(q, J=7.6 Hz, 2H), 2.43 (t, J=7.2 Hz, 2H), 1.71 (quint, J=7.2 Hz, 2H),1.10 (t, J=7.6 Hz, 3H).

Step 2:

(E/Z)-2-(3-(3-(2-Ethylstyryl)phenyl)propyl)isoindoline-1,3-dione wasdeprotected to afford Example 36 as an oil. Yield (0.1263 g, 45%),trans-/cis-isomer ratio 1:3. Cis-isomer: ¹H NMR (400 MHz, DMSO-d₆) δ6.86-7.31 (m, 8H), 6.74 (d, J=12.0 Hz, 1H), 6.63 (d, J=12.4 Hz, 1H),2.63 (t, J=7.2 Hz, 2H), 2.57-2.59 (m, 2H), 2.41 (t, J=7.2 Hz, 2H), 1.45(quint, J=7.2 Hz, 2H), 1.12 (t, J=7.6 Hz, 3H).

Example 37 Preparation of(E/Z)-3-(3-(2-ethynylstyryl)phenyl)propan-1-amine

(E/Z)-3-(3-(2-Ethynylstyryl)phenyl)propan-1-amine was prepared accordingto the method used in Example 32.

Step 1:

2-Ethynylbenzyltriphenylphosphonium bromide was prepared from2-ethynylbenzyl bromide to give a white solid. Yield (0.599 g, 60%): ¹HNMR (400 MHz, DMSO-d₆) δ 7.71-7.98 (m, 19H), 5.18 (d, J=12.0 Hz, 2H),4.20 (s, 1H).

Step 2:

Phthalimide 29 was coupled with 2-ethynyllbenzyltriphenylphosphoniumbromide and purified by flash chromatography (10 to 50% EtOAc-hexanesgradient) to give(E)-2-(3-(3-(2-ethynylstyryl)phenyl)propyl)isoindoline-1,3-dione. Yield(0.3055 g, 78%), trans-/cis-isomer ratio 1.3:1. Cis-isomer: ¹H NMR (400MHz, DMSO-d₆) δ 7.80-7.87 (m, 4H), 7.40-7.54 (m, 1H), 7.35 (t, J=7.6 Hz,1H), 7.29 (dt, J=7.6, 2.4 Hz, 2H), 6.99-7.21 (m, 3H), 6.92 (d, J=7.6 Hz,1H), 6.75 (d, J=12.0 Hz, 1H), 6.69 (d, J=12.0 Hz, 1H), 4.41 (s, 1H),3.52 (t, J=7.2 Hz, 2H), 2.47-2.51 (m, 2H), 1.76 (quint, J=7.6 Hz, 2H).

Step 3:

(E)-2-(3-(3-(2-ethynylstyryl)phenyl)propyl)isoindoline-1,3-dione wasdeprotected to afford Example 37 as an oil. Yield (0.0417 g, 20%),trans-/cis-isomer ratio 1:2. Cis-isomer: ¹H NMR (400 MHz, MeOD) δ7.02-7.53 (m, 8H), 6.84 (d, J=12.0 Hz, 1H), 6.71 (d, J=12.0 Hz, 1H),3.76 (s, 1H), 2.86 (t, J=8.0 Hz, 2H), 2.59 (t, J=8.0 Hz, 2H), 1.87(quint, J=8.0, 2H).

Example 38 Preparation of(E/Z)-3-(3-(3,4-dimethylstyryl)phenyl)propan-1-amine

(E/Z)-3-(3-(3,4-Dimethylstyryl)phenyl)propan-1-amine was preparedaccording to the method used in Example 32.

Step 1:

3,4-Dimethylbenzyltriphenylphosphonium bromide was coupled withphthalimide 29 to give(E/Z)-2-(3-(3-(3,4-dimethylstyryl)phenyl)propyl)isoindoline-1,3-dione asan oil. Yield (0.2311 g, 42%), trans-/cis-isomer ratio ˜1:1. Cis-isomer:¹H NMR (400 MHz, CDCl₃) δ 7.69-7.76 (m, 4H), 6.96-7.35 (m, 7H), 6.54 (d,J=12.8 Hz, 1H), 6.50 (d, J=12.4 Hz, 1H), 3.80 (t, J=7.6 Hz, 2H), 2.74(t, J=7.6 Hz, 2H), 2.32 (s, 3H), 2.30 (s, 3H), 2.11 (quint, J=7.6 Hz,2H).

Step 2:

(E/Z)-2-(3-(3-(3,4-Dimethylstyryl)phenyl)propyl)isoindoline-1,3-dionewas deprotected to give Example 38 as an oil. Trans-/cis-isomer ratio1:1.7. Cis-isomer: ¹H NMR (400 MHz, DMSO-d₆) δ 7.32-7.39 (m, 3H), 7.26(t, J=8.0 Hz, 1H), 6.85-7.22 (m, 5H), 2.78-2.86 (m, 2H), 2.69 (t, J=7.6Hz, 2H), 2.30 (s, 3H), 2.27 (s, 3H), 1.85 (quint, J=7.6 Hz, 2H).

Example 39 Preparation of(E/Z)-3-(3-(2-isopropylstyryl)phenyl)propan-1-amine

(E/Z)-3-(3-(2-Isopropylstyryl)phenyl)propan-1-amine was preparedaccording to the method used in Example 32.

Step 1:

2-Isopropyllbenzyltriphenylphosphonium bromide was prepared from2-isopropylbenzyl bromide according to the method used in Example 32,except that the reaction was heated for 2 h. The product was isolated asa white solid. Yield (1.05 g, 82%): ¹H NMR (400 MHz, DMSO-d₆) δ7.71-7.72 (m, 3H), 7.60-7.62 (m, 6H), 7.57-7.60 (m, 6H), 7.24-7.29 (m,2H), 6.99 (t, J=7.0 Hz, 1H), 6.87 (dq, J=7.6, 1.2 Hz, 1H), 5.01 (d,J=5.1 Hz, 2H), 2.55-2.62 (m, 1H), 0.73 (d, J=6.8 Hz, 6H).

Step 2:

Phthalimide 29 was coupled with 2-isopropyllbenzyltriphenylphosphoniumbromide according to the method used in Example 32, except that thereaction was not warmed to room temperature. Purification by flashchromatography (10 to 50% EtOAc-hexanes gradient) gave(E)-2-(3-(3-(2-isopropylstyryl)phenyl)propyl)isoindoline-1,3-dione as anoil. Yield (0.4283 g, 73%), trans-/cis-isomer ratio ˜1:1. Trans-isomer:¹H NMR (400 MHz, DMSO-d₆) δ 7.78-7.84 (m, 4H), 7.56 (d, J=8.0 Hz, 1H),7.51 (d, J=16.0 Hz, 1H), 7.46 (s, 1H), 6.80-7.30 (m, 7H), 3.47 (t, J=6.8Hz, 2H), 3.08-3.14 (m, 1H), 2.39 (t, J=6.8 Hz, 2H), 1.61-1.68 (m, 2H),1.09 (d, J=6.8 Hz, 6H).

Step 3:

(E)-2-(3-(3-(2-Isopropylstyryl)phenyl)propyl)isoindoline-1,3-dione wasdeprotected according to the method used in Example 32 except that thereaction was heated to reflux for 1.3 h. Example 39 was isolated as anoil. Yield (0.1838 g, 63%), trans-/cis-isomer ratio 1:3. Cis-isomer: ¹HNMR (400 MHz, DMSO-d₆) δ 7.18-7.34 (m, 2H), 7.01-7.10 (m, 2H), 6.92-6.98(m, 2H), 6.81-6.84 (m, 2H), 6.77 (d, J=12.0 Hz, 1H), 6.61 (d, J=12.0 Hz,1H), 3.08-3.15 (m, 1H), 2.32-2.48 (m, 4H), 1.36-1.42 (m, 2H), 1.24 (brs, 2H), 1.12 (d, J=6.8 Hz, 6H).

Example 40 Preparation of(E/Z)-4-(3-(3,5-dimethylstyryl)phenyl)propan-1-amine

(E/Z)-4-(3-(3,5-Dimethylstyryl)phenyl)propan-1-amine was preparedaccording to the method used in Example 32.

Step 1:

3,5-Dimethylbenzyltriphenylphosphonium bromide was coupled withphthalimide 29. Purification by flash chromatography (10 to 50%EtOAc-hexanes gradient) gave(E)-2-(3-(3-(3,5-dimethylstyryl)phenyl)propyl)isoindoline-1,3-dione as awhite solid. Yield (0.3263 g, 55%): ¹H NMR (400 MHz, DMSO-d₆) δ7.81-7.88 (m, 4H), 7.00-7.44 (m, 6H), 6.71-6.90 (m, 2H), 6.53 (s, 1H),3.64 (t, J=7.2 Hz, 2H), 2.65 (t, J=7.2 Hz, 2H), 1.70-1.77 (m, 2H), 2.29(s, 6H).

Step 2:

(E)-2-(3-(3-(3,5-Dimethylstyryl)phenyl)propyl)isoindoline-1,3-dione wasdeprotected to give Example 40 as an oil. Yield (0.1632 g, 82%),trans-/cis-isomer ratio 1:1.3. Cis-isomer: ¹H NMR (400 MHz, DMSO-d₆) δ7.02-7.40 (m, 5H), 6.82-6.89 (m, 2H), 6.54 (d, J=12.4 Hz, 1H), 6.50 (d,J=12.4 Hz, 1H), 2.54 (t, J=6.8 Hz, 2H), 2.43-2.46 (m, 2H), 2.14 (s, 6H),1.46-1.54 (m, 2H), 1.30 (br s, 2H).

Example 41 Preparation of(E/Z)-4-(3-(2-methoxystyryl)phenyl)propan-1-amine

(E/Z)-4-(3-(2-Methoxystyryl)phenyl)propan-1-amine was prepared accordingto the method used in Example 32.

Step 1:

2-Methoxybenzyltriphenylphosphonium bromide was coupled with phthalimide29 to give(E)-2-(3-(3-(2-methoxystyryl)phenyl)propyl)isoindoline-1,3-dione as awhite solid. Yield (0.5590 g, quant.), trans-/cis-isomer ratio 1:1.Trans-isomer: ¹H NMR (400 MHz, DMSO-d₆) δ 7.81-7.88 (m, 4H), 7.52-7.65(m, 5H), 6.69-7.41 (m, 4H), 6.57 (d, J=16.8 Hz, 1H), 3.76 (s, 3H), 3.52(t, J=6.8 Hz, 2H), 2.48 (t, J=8.0 Hz, 2H), 1.91-1.99 (m, 2H).

Step 2:

(E)-2-(3-(3-(2-methoxystyryl)phenyl)propyl)isoindoline-1,3-dione wasdeprotected to give Example 41 as an oil. Yield (0.260 g, 73%),trans-/cis-isomer ratio 1:1. Trans-isomer: ¹H NMR (400 MHz, DMSO-d₆) δ6.90-7.65 (m, 9H), 6.57 (d, J=16.8 Hz, 1H), 3.78 (s, 3H), 2.52 (t, J=6.8Hz, 2H), 2.41-2.46 (m, 2H), 1.22-1.26 (m, 2H), 1.14 (br s, 2H).

Example 42 Preparation of(E)-2-(3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propylamino)ethanol

(E)-2-(3-(3-(2-(2,6,6-Trimethylcyclohex-1-enyl)vinyl)phenyl)propylamino)ethanolwas prepared according to Scheme 16.

Step 1:

Iodide 31 was reacted with allyl alcohol following the method used inExample 32. Following the reaction, the mixture was partitioned betweenEtOAc and water. The combined organics were washed with water and brine,dried over Na₂SO₄ and concentrated under reduced pressure. Purificationby flash chromatography (0 to 20% EtOAc-hexanes gradient) gave aldehyde52 as a yellow oil. Yield (0.375 g, 63%).

Step 2:

To a solution of aldehyde 52 (0.325 g, 1.15 mmol) in MeOH (2 mL) wasadded ethanolamine (0.08 g, 1.4 mmol) and 4 Å molecular sieves. Themixture was stirred for 2 h, then NaBH₄ (0.068 g, 1.8 mmol) was addedand the reaction was stirred overnight. Solids were removed byfiltration and the filtrate was concentrated under reduced pressure. Theresidue was partitioned between EtOAc and water then the combinedorganics were washed with brine, dried over Na₂SO₄ and concentratedunder reduced pressure. Purification was conducted by flashchromatography (0-10% 7 M NH₃-MeOH in EtOAc) three times. HPLCpurification (30 to 90% MeCN—H₂O gradient) gave Example 42 as an oil.Yield (0.033 g, 9%), trans-/cis-isomer ratio 9:1. Trans-isomer: ¹H NMR(400 MHz, CDCl₃) δ 7.21-7.26 (m, 3H), 7.03-7.05 (m, 1H), 6.66 (d, J=16.4Hz, 1H), 6.31 (d, J=16.4 Hz, 1H), 3.56 (t, J=5.1 Hz, 2H), 2.69 (t, J=5.2Hz, 2H), 2.57-2.62 (m, 4H), 2.55 (br s, 2H), 1.96 (t, J=6.1 Hz, 2H),1.72-1.80 (m, 2H), 1.76 (s, 3H), 1.54-1.60 (m, 2H), 1.40-1.47 (m, 2H),1.06 (s, 6H).

Example 43 Preparation of(E)-3-(3-(2,6-dichlorostyryl)phenyl)propan-1-amine

(E)-3-(3-(2,6-Dichlorostyryl)phenyl)propan-1-amine was preparedaccording to the method used in Example 32.

Step 1:

2,6-Dichlorobenzyltriphenylphosphonium bromide was coupled withphthalimide 29. Purification by flash chromatography (10 to 50%EtOAc-hexanes gradient) gave(E)-2-(3-(3-(2,6-dichlorostyryl)phenyl)propyl)isoindoline-1,3-dione as awhite solid. Yield (0.7041 g, 96%): ¹H NMR (400 MHz, DMSO-d₆) δ7.78-7.86 (m, 4H), 7.51 (d, J=8.4 Hz, 2H), 7.44 (s, 1H), 7.37 (d, J=7.6Hz, 1H), 7.31 (d, J=8.4 Hz, 1H), 7.27 (t, J=7.2 Hz, 1H), 7.18 (d, J=7.6Hz, 1H), 7.11 (d, J=16.4 Hz, 1H), 7.01 (d, J=16.8 Hz, 1H), 3.62 (t,J=6.8 Hz, 2H), 2.65 (t, J=7.4 Hz, 2H), 1.88-1.98 (m, 2H).

Step 2:

(E)-2-(3-(3-(2,6-dichlorostyryl)phenyl)propyl)isoindoline-1,3-dione wasdeprotected and purified by flash chromatography (1:5:5 7 M NH₃-MeOH:EtOAc: heptane) to give Example 43 as an oil. Yield (0.1123 g, 23%): ¹HNMR (400 MHz, DMSO-d₆) δ 7.51 (d, J=8.2 Hz, 2H), 7.40-7.42 (m, 2H),7.27-7.33 (m, 2H), 7.16 (d, J=7.2 Hz, 1H), 7.12 (d, J=16.8 Hz, 1H), 7.03(d, J=16.6 Hz, 1H), 2.61 (t, J=7.8 Hz, 2H), 2.53 (t, J=7.9 Hz, 2H),1.61-1.72 (m, 2H), 1.40 (br s, 2H).

Example 44 Preparation of(E/Z)-3-(3-(2,3-dimethylstyryl)phenyl)propan-1-amine

(E/Z)-3-(3-(2,3-Dimethylstyryl)phenyl)propan-1-amine was preparedaccording to the method used in Example 32.

Step 1:

To an ice cold solution of 2,3-dimethylbenzyltriphenylphosphoniumbromide (1.1197 g, 2.43 mmol) in CH₂Cl₂ (10 mL) was added a solution ofpotassium tert-butoxide (2.5 mL of a1 M solution in THF, 2.5 mmol) Themixture was stirred for 5 min, then a solution of phthalimide 29 (0.3380g, 1.15 mmol) in CH₂Cl₂ (10 mL) was added. The mixture was warmed toroom temperature, stirred for 20 min then concentrated under reducedpressure. 10% EtOAc-heptane was added and the solids were removed byfiltration. The filtrate was concentrated under reduced pressure.Purification by repeated flash chromatography (6 to 40% EtOAc-hexanesgradient then 10% EtOAc-hexanes) gave(Z)-2-(3-(3-(2,3-dimethylstyryl)phenyl)propyl)isoindoline-1,3-dione asan oil. Yield (0.1871 g, 41%), trans-/cis-isomer ratio 1:11.7.Cis-isomer: ¹H NMR (400 MHz, CDCl₃) δ 7.85 (dd, J=5.6, 2.8 Hz, 2H), 7.72(dd, J=5.2, 3.2 Hz, 2H), 7.04 (t, J=7.6 Hz, 1H), 6.88-6.98 (m, 6H), 6.68(d, J=12.0 Hz, 1H), 6.56 (d, J=12.0 Hz, 1H), 3.64 (t, J=7.2 Hz, 2H),2.52 (t, J=7.6 Hz, 2H), 2.52 (s, 3H), 2.17 (s, 3H), 1.80-1.88 (m, 2H).

Step 2:

(Z)-2-(3-(3-(2,3-Dimethylstyryl)phenyl)propyl)isoindoline-1,3-dione wasdeprotected and purified according to the method used in Example 32 togive Example 43 as an oil. Yield (0.0662 g, 54%), trans-/cis-isomerratio 1:4. Cis-isomer: ¹H NMR (400 MHz, DMSO-d₆) δ 6.82-7.07 (m, 7H),6.68 (d, J=12.0 Hz, 1H), 6.59 (d, J=12.0 Hz, 1H), 2.41 (t, J=7.6 Hz,4H), 2.25 (s, 3H), 2.14 (s, 3H), 1.40-1.46 (m, 2H), 1.32 (br s, 2H).

Example 45 Preparation of(E/Z)-3-(3-(2,6-dimethylstyryl)phenyl)prop-2-en-1-amine

(E/Z)-3-(3-(2,6-Dimethylstyryl)phenyl)prop-2-en-1-amine (isomer ratio80:20 trans: cis) was prepared according to Scheme 17.

Step 1:

To a solution of 2,6-dimethylbenzylphosphonium bromide (3.46 g, 7.5mmol) in CH₂Cl₂ (20 mL) was added a solution of potassium tert-butoxide(7.5 mL of a 1 M solution in THF, 7.5 mmol) The mixture was stirred atroom temperature for 10 min, then 3-bromobenzaldehyde (0.925 g, 5 mmol)was added and the mixture stirred for 4 h. The reaction mixture wasconcentrated under reduced pressure and partitioned between EtOAc andwater then solids were removed by filtration. The combined organics werewashed with brine, dried over MgSO₄ and concentrated under reducedpressure. Purification by flash chromatography (7 to 60% EtOAc-hexanesgradient) gave aryl bromide 53 as a colorless oil. Yield (0.930 g, 65%),trans-/cis-isomer ratio 10:1.

trans-isomer: ¹H NMR (400 MHz, DMSO-d₆) δ 7.81 (t, J=1.6 Hz, 1H),7.59-7.61 (m, 1H), 7.44-7.46 (m, 1H), 7.27-7.34 (m, 2H), 7.04-7.14 (m,3H), 6.64 (d, J=16.8 Hz, 1H), 2.30 (s, 6H).

Step 2:

To a solution of aryl bromide 53 (0.343 g, 1.2 mmol) and tert-butylallylcarbamate (0.189 g, 1.2 mmol) in triethylamine (0.33 mL, 2.4 mmol)and acetonitrile (5 mL) was added Pd(OAc)₂ (0.014 g, 0.06 mmol) andtri-o-tolylphosphine (0.018 g, 0.06 mmol). The mixture was bubbled withargon then heated to 70° C. for 6 h. After cooling to room temperature,the mixture was concentrated under reduced pressure and partitionedbetween EtOAc and water. The combined organics were washed with brine,treated with activated charcoal and dried over a mixture of MgSO₄ andNa₂SO₄. After concentration under reduced pressure, purification byflash chromatography (2 to 20% EtOAc-hexanes gradient) gave impure allylamine 54 as an oil. Yield (0.059 g, contaminated with ˜50% tert-butylallylcarbamate; 10% yield): ¹H NMR (400 MHz, DMSO-d₆) δ 7.45 (d, J=6.4Hz, 1H), 7.28-7.34 (m, 2H), 7.24 (d, J=16.8 Hz, 1H), 7.03-7.10 (m, 5H),6.64 (d, J=16.8 Hz, 1H), 6.46 (d, J=16.0 Hz, 1H), 6.27 (dt, J=15.6, 5.6Hz, 1H), 3.72 (t, J=5.6 Hz, 2H), 2.31 (s, 6H), 1.36 (s, 9H).

Step 3:

To a solution of allyl amine 54 (0.059 g, impure; 0.11 mmol) in diethylether (2 mL) was added a solution of HCl-diethyl ether (3 mL of ˜10 Msolution, 30 mmol) The mixture was stirred at room temperature for 1 hthen concentrated under reduced pressure. To the residue was added 7 MNH₃-MeOH (5 mL) and the solution was concentrated under reducedpressure. Purification by flash chromatography (15% 7 M NH₃ inMeOH-EtOAc) gave Example 45 (80% all-trans) as a colorless oil. Yield(0.0202 g, 70%). Trans-isomer: ¹H NMR (400 MHz, DMSO-d₆) δ 7.59 (br s,1H), 7.41-7.45 (m, 1H), 7.29-7.31 (m, 2H), 7.23 (d, J=16.9 Hz, 1H),7.01-7.15 (m, 3H), 6.64 (d, J=16.8 Hz, 1H), 6.49-6.53 (m, 1H), 6.43 (t,J=4.8 Hz, 1H), 3.32 (dd, J=5.2, 1.2 Hz, 2H), 2.31 (s, 6H).

Example 46 Preparation of(E)-3-(3-(2,6-dimethylstyryl)-4-fluorophenyl)propan-1-amine

(E)-3-(3-(2,6-Dimethylstyryl)-4-fluorophenyl)propan-1-amine was preparedaccording to the method used in Example 1 with modifications byreplacing 3-iodobenzaldehyde with 6-fluoro-3-iodobenzaldehyde in Step 3.

Step 1:

To a solution of 2,6-dimethylbenzyltriphenylphosphonium bromide (2.21 g,4.8 mmol) in CH₂Cl₂ (30 mL) was added a solution of potassiumtert-butoxide (5 mL of a 1 M solution in THF, 5 mmol) and the mixturewas stirred at 0° C. for 15 min. The mixture was cooled to −78° C. and6-fluoro-3-iodobenzaldehyde (1.0 g, 4.0 mmol) was added. The reactionmixture was allowed to warm to room temperature and stirred overnight.The mixture was partitioned between EtOAc and water and the combinedorganics were washed with 1 M HCl and brine, dried over Na₂SO₄ andconcentrated under reduced pressure. Purification by flashchromatography (100% hexanes) gave(E)-2-(2-fluoro-5-iodostyryl)-1,3-dimethylbenzene (0.280 g) and(Z)-2-(2-fluoro-5-iodostyryl)-1,3-dimethylbenzene (0.472 g) withadditional product collected as an E/Z mixture (0.450 g). Total yield(1.20 g, 85%). Trans-isomer: ¹H NMR (400 MHz, DMSO-d₆) δ 8.12 (dd,J=7.2, 2.4 Hz, 1H), 7.63 (dq, J=8.8, 2.4 Hz, 1H), 7.34 (d, J=16.8 Hz,1H), 7.04-7.09 (m, 4H), 6.61 (d, J=16.8 Hz, 1H), 2.30 (s, 6H).

Step 2:

(E)-2-(2-Fluoro-5-iodostyryl)-1,3-dimethylbenzene was coupled with allylalcohol according to the method in Example 42. Purification by flashchromatography (40% CH₂Cl₂-hexanes) gave(E)-3-(3-(2,6-dimethylstyryl)-4-fluorophenyl)propanal as an oil. Yield(0.115 g, 52%): ¹H NMR (400 MHz, DMSO-d₆) δ 9.72 (t, J=1.2 Hz, 1H), 7.64(dd, J=7.2, 2.4 Hz, 1H), 7.27 (d, J=16.8 Hz, 1H), 7.09-7.19 (m, 2H),7.06 (s, 3H), 6.67 (d, J=16.8 Hz, 1H), 2.78-2.89 (m, 4H), 2.31 (s, 6H).

Step 3:

To a solution of (E)-3-(3-(2,6-dimethylstyryl)-4-fluorophenyl)propanal(0.110 g, 0.39 mmol) in anhydrous MeOH (5 mL) was added a solution ofNH₃-MeOH (10 mL of a 7 M solution). The reaction mixture was stored at−5° C. overnight, then warmed to room temperature. NaBH₄ (0.148 g, 3.9mmol) was added and the mixture was stirred at room temperature for 2 h.After concentration under reduced pressure, the residue was partitionedbetween saturated aqueous NaHCO₃ and EtOAc. The combined organics weredried over Na₂SO₄ and concentrated under reduced pressure. Purificationby flash chromatography (5 to 10% 7 M NH₃-MeOH in CH₂Cl₂) gave Example46 as an oil. (Yield 0.022 g, 20%): ¹H NMR (400 MHz, CDCl₃) δ 7.40 (dd,J=7.2, 2.4 Hz, 1H), 7.17 (d, J=16.8 Hz, 1H), 7.04-7.10 (m, 4H), 6.99(dd, J=10.4, 8.4 Hz, 1H), 6.74 (d, J=16.8 Hz, 1H), 2.78 (t, J=7.2 Hz,2H), 2.68 (t, J=8.0 Hz, 2H), 2.39 (s, 6H), 1.77-1.85 (m, 2H).

Example 47 Preparation of(E/Z)-3-(3-(2-(trifluoromethyl)styryl)phenyl)propan-1-amine

(E/Z)-3-(3-(2-(Trifluoromethyl)styryl)phenyl)propan-1-amine was preparedaccording to the method used in Example 32.

Step 1:

2-Trifluoromethylbenzyltriphenylphosphonium bromide was coupled withphthalimide 29 following the method used in Example 44 except that thereaction was conducted at −78° C. instead of 0° C. before warming toroom temperature. Purification by flash chromatography (10 to 40%EtOAc-hexanes gradient) gave(E)-2-(3-(3-(2-trifluoromethylstyryl)phenyl)propyl)isoindoline-1,3-dione(0.0797 g, trans-/cis-isomer ratio 15.6:1) as an oil and(Z)-2-(3-(3-(2-trifluoromethylstyryl)phenyl)propyl)isoindoline-1,3-dione(0.1139 g, trans-/cis-isomer ratio 8.1:1) as an oil. Yield (0.1936 gtotal, 83%): Trans-isomer: ¹H NMR (400 MHz, CDCl₃) δ 7.69-7.73 (m, 2H),7.68 (d, J=7.6 Hz, 1H), 7.56-7.63 (m, 3H), 7.45 (t, J=7.6 Hz, 1H), 7.34(dq, J=16.0, 2.0 Hz, 1H), 7.21-7.28 (m, 3H), 7.14-7.19 (m, 1H), 7.05 (d,J=7.6 Hz, 1H), 6.93 (d, J=16.4 Hz, 1H), 3.69 (t, J=7.2 Hz, 2H), 2.64 (t,J=7.2 Hz, 2H), 1.95-2.04 (m, 2H).

Step 2:

(E)-2-(3-(3-(2-Trifluorostyryl)phenyl)propyl)isoindoline-1,3-dione wasdeprotected according to the method used in Example 32, except that thereaction was heated at 60° C. overnight. Purification by flashchromatography (10% 7 M NH₃ in MeOH-EtOAc) gave Example 47 as an oil.Yield (0.0180 g, 32%), trans-/cis-isomer ratio 5.2:1. Trans-isomer: ¹HNMR (400 MHz, CDCl₃) δ 7.77 (d, J=8.0 Hz, 1H), 7.65 (t, J=8.0 Hz, 1H),7.54 (t, J=7.6 Hz, 1H), 7.46 (dq, J=16.4, 1.2 Hz, 1H), 7.26-7.38 (m,4H), 7.14 (d, J=7.2 Hz, 1H), 7.06 (d, J=16.4 Hz, 1H), 2.68-2.78 (m, 4H),1.72-1.84 (m, 2H), 1.20 (br s, 2H).

Example 48 Preparation of(E)-3-(3-(2,6-dimethoxystyryl)phenyl)propan-1-amine

(E)-3-(3-(2,6-dimethoxystyryl)phenyl)propan-1-amine was preparedaccording to the method used in Example 32.

Step 1:

2,6-Dimethoxybenzyltriphenylphosphonium bromide was coupled withphthalimide 29 following the method used in Example 44, except that thereaction was stirred overnight after warming up to room temperature.After concentration under reduced pressure, the residue was partitionedbetween EtOAc and water. The combined organics were washed with brine,dried over MgSO₄ and concentrated under reduced pressure. Purificationby flash chromatography (7 to 60% EtOAc-hexanes gradient) gave(E)-2-(3-(3-(2,6-dimethoxystyryl)phenyl)propyl)isoindoline-1,3-dione asa light yellow oil. Yield (0.317 g, 82%): ¹H NMR (400 MHz, DMSO-d₆) δ7.79-7.84 (m, 4H), 7.42 (d, J=17.2 Hz, 1H), 7.17-7.32 (m, 5H), 7.06-7.09(m, 1H), 6.58 (d, J=8.4 Hz, 2H), 3.83 (s, 6H), 3.61 (t, J=7.2 Hz, 2H),2.63 (t, J=8.0 Hz, 2H), 1.88-1.96 (m, 2H).

Step 2:

(E)-2-(3-(3-(2,6-dimethoxystyryl)phenyl)propyl)isoindoline-1,3-dione wasdeprotected according to the method used in Example 32 except that thereaction was conducted overnight at room temperature in MeOH. Afterconcentration under reduced pressure, the residue was suspended in EtOAcand sonicated then solids were removed by filtration. The filtrate wasconcentrated under reduced pressure. Purification by flashchromatography (0.3% concentrated aqueous NH₄OH/10% 7 M NH₃-MeOH/90%EtOAc) gave Example 48 as a colorless oil. Yield (0.179 g, 83%): ¹H NMR(400 MHz, DMSO-d₆) δ 7.44 (d, J=16.8 Hz, 1H), 7.31 (d, J=16.8 Hz, 1H),7.27 (s, 2H), 7.25 (d, J=7.6 Hz, 1H), 7.19 (t, J=8.4 Hz, 1H), 7.05 (d,J=7.2 Hz, 1H), 6.68 (d, J=8.4 Hz, 2H), 3.83 (s, 6H), 2.59 (t, J=7.2 Hz,2H), 2.53 (t, J=8.0 Hz, 2H), 1.60-1.67 (m, 2H), 1.35 (br s, 2H).

Example 49 Preparation of(E)-3-(3-(2,6-bis(trifluoromethyl)styryl)phenyl)propan-1-amine

(E)-3-(3-(2,6-Bis(trifluoromethyl)styryl)phenyl)propan-1-amine wasprepared according to the method used in Example 32.

Step 1:

To a solution of 2,6-bis(trifluoromethyl)benzyl bromide (0.400 g, 1.30mmol) in toluene (5 mL) was added triphenylphosphine (0.375 g, 1.43mmol) The mixture was heated to reflux for 24 h. After cooling to roomtemperature, hexanes (10 mL) was added and the mixture was stirred for 1h. The solid was collected by filtration and rinsed with hexanes to give2,6-bis(trifluoromethyl)benzyltriphenylphosphonium bromide as a whitesolid. Yield (0.446 g, 60%): ¹H NMR (400 MHz, DMSO-d₆) δ 8.02 (d, J=8.0Hz, 2H), 7.57-7.87 (m, 16H), 5.17 (d, J=15.2 Hz, 2H).

Step 2:

2,6-Bis(trifluoromethyl)benzyltriphenylphosphonium bromide was coupledwith phthalimide 29 following the method used in Example 46.Purification by flash chromatography (30% diethyl ether-hexanes) gave(E)-2-(3-(3-(2,6-bis(trifluoromethyl)styryl)phenyl)propyl)isoindoline-1,3-dioneas an oil. Yield (0.227 g, 62%): ¹H NMR (400 MHz, DMSO-d₆) δ 8.09 (d,J=8.0 Hz, 2H), 7.78-7.84 (m, 4H), 7.75 (t, J=7.6 Hz, 1H), 7.28-7.38 (m,3H), 7.27 (t, J=7.6 Hz, 1H), 7.19 (d, J=7.2 Hz, 1H), 6.57 (d, J=16.8 Hz,1H), 3.62 (t, J=6.8 Hz, 2H), 2.64 (t, J=8.0 Hz, 2H), 1.88-1.96 (m, 2H).

Step 3:

(E)-2-(3-(3-(2,6-Bis(trifluoromethyl)styryl)phenyl)propyl)isoindoline-1,3-dionewas deprotected according to the method used in Example 48. Afterconcentration under reduced pressure, the residue was suspended indiethyl ether and sonicated. Solids were removed by filtration and thefiltrate was concentrated under reduced pressure. Purification by flashchromatography (10% 7 M NH₃ in MeOH/CH₂Cl₂) gave Example 49 as an oil.Yield (0.102 g, 63%): ¹H NMR (400 MHz, DMSO-d₆) δ 8.09 (d, J=8.0 Hz,2H), 7.75 (t, J=8.4 Hz, 1H), 7.27-7.38 (m, 4H), 7.16 (d, J=7.6 Hz, 1H),6.59 (d, J=16.8 Hz, 1H), 2.61 (t, J=6.8 Hz, 2H), 2.55 (t, J=8.0 Hz, 2H),1.88 (br s, 2H), 1.61-1.68 (m, 2H).

Example 50 Preparation of(E)-3-amino-1-(3-(2,6-dichlorostyryl)phenyl)propan-1-ol

(E)-3-Amino-1-(3-(2,6-dichlorostyryl)phenyl)propan-1-ol was preparedaccording to Scheme 18.

Step 1:

2,6-Dichlorobenzyltriphenylphosphonium bromide (55) was coupled with3-bromobenzaldehyde (7) according to the method in Example 46 exceptthat the reaction was warmed to room temperature briefly after theaddition of potassium tert-butoxide. It was cooled to −78° C. againbefore the addition of 3-bromobenzaldehyde. After warming to roomtemperature and stirring overnight, the reaction mixture wasconcentrated under reduced pressure. The residue was triturated with ˜5%EtOAc-hexanes and solids were removed by filtration. The filtrate wasconcentrated under reduced pressure and the residue was recrystallizedfrom ˜2-5% EtOAc-hexanes to give bromide 56 as a white crystallinesolid. Yield (1.666 g, 78%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.84 (t, J=1.8Hz, 1H), 7.59-7.65 (m, 2H), 7.50-7.59 (m, 2H), 7.31-7.37 (m, 2H), 7.21(d, J=16.8 Hz, 1H), 7.08 (d, J=16.8 Hz, 1H).

Step 2:

To a −78° C. solution of bromide 56 (0.7008 g, 2.14 mmol) in THF (10 mL)was added n-butyl lithium (1.1 mL of a 2.5 M solution in THF, 2.75 mmol)and the mixture was stirred for 9 min. DMF (0.3 mL, 3.9 mmol) was addedthen the reaction mixture was stirred for 10 min. After allowing to warmto room temperature, additional DMF (0.3 mL, 3.9 mmol) was added and thereaction was stirred for 22 min. The mixture was partitioned betweenbrine and EtOAc. The combined organics were washed with brine, driedover MgSO₄ and concentrated under reduced pressure. Purification byflash chromatography (10 to 40% EtOAc-hexanes gradient) gave aldehyde 57as a white solid. Yield (0.359 g, 61%): ¹H NMR (400 MHz, CDCl₃) δ 10.07(s, 1H), 8.04 (t, J=1.6 Hz, 1H), 7.79-7.83 (m, 2H), 7.56 (t, J=7.6 Hz,1H), 7.37 (d, J=7.6 Hz, 2H), 7.23 (d, J=8.0 Hz, 2H), 7.14 (t, J=8.4 Hz,1H).

Step 3:

The glassware used in this synthetic transformation was dried with aheat gun under vacuum. To a −78° C. solution of lithium diisopropylamide(1.0 mL of a 2 M solution in THF, 2.0 mmol) in THF (3 mL) was added asolution of acetonitrile (0.1 mL, 1.88 mmol) in THF (5 mL) slowly via anaddition funnel. The reaction mixture was stirred at −78° C. for 12 min,then a solution of aldehyde 57 (0.353 g, 1.27 mmol) in THF (6 mL) wasadded dropwise over 12 min. Additional THF (3 mL) was added and themixture was stirred for 25 min. After warming to room temperature, thereaction was quenched with brine and extracted with EtOAc. The combinedorganics were washed with brine, dried over MgSO₄ and concentrated underreduced pressure. Purification by flash chromatography (10 to 70%EtOAc-hexanes gradient) gave nitrile 58 as a white solid. Yield (0.357g, 88%): ¹H NMR (400 MHz, CDCl₃) δ 7.55 (d, J=1.2 Hz, 1H), 7.53 (t,J=1.2 Hz, 1H), 7.40-7.44 (m, 1H), 7.34-7.37 (m, 3H), 7.10-7.15 (m, 3H),5.07-5.12 (m, 1H), 2.81 (dd, J=6.8, 2.0 Hz, 2H), 2.44 (d, J=3.6 Hz, 1H).

Step 4:

To an ice cold solution of nitrile 58 (0.357 g, 1.12 mmol) in THF (7 mL)was added a solution of LiAlH₄ (1 mL of a 2 M solution in THF, 2.0mmol). The reaction mixture was stirred for 30 min at 0° C. thenquenched with saturated aqueous Na₂SO₄. The solution was dried overMgSO₄ and concentrated under reduced pressure. Purification by flashchromatography (9:9:2 EtOAc:heptane:7 M NH₃-MeOH) gave Example 50 as anoil. Yield (0.0785 g, 22%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.51-7.54 (s,1H), 7.52 (d, J=8.0 Hz, 2H), 7.45 (d, J=7.6 Hz, 1H), 7.27-7.35 (m, 3H),7.12 (d, J=16.8 Hz, 1H), 7.06 (d, J=16.8 Hz, 1H), 4.69 (t, J=7.2 Hz,1H), 2.60-2.70 (m, 2H), 1.63-1.68 (m, 2H).

Example 51 Preparation of(E/Z)—N-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)acetamide

(E/Z)—N-(3-(3-(2,6-Dimethylstyryl)phenyl)propyl)acetamide (isomer ratio80:20 trans:cis) was prepared according to Scheme 19.

Step 1:

To a solution of (E/Z)-3-(3-(2,6-dimethylstyryl)phenyl)propan-1-amine(isomer ratio 80:20 trans:cis, Example 1) (0.049 g, 0.185 mmol) inCH₂Cl₂ (2 mL) was added triethylamine (0.065 mL, 0.47 mmol), aceticanhydride (0.020 mL, 0.21 mmol) and N, N-dimethylaminopyridine (˜5 mg).The reaction mixture was stirred at room temperature for 1.25 h. Themixture was concentrated under reduced pressure and the residue waspartitioned between EtOAc and water. The combined organics were washedwith saturated aqueous NaHCO₃ and brine, dried over Na₂SO₄ andconcentrated under reduced pressure. Purification by flashchromatography (50 to 100% EtOAc-hexanes gradient) gave Example 51 as acolorless oil. Yield (0.037 g, 65%), trans-/cis-isomer 5:1. Transisomer: ¹H NMR (400 MHz, CDCl₃) δ 7.29-7.35 (m, 3H), 7.08-7.11 (m, 5H),6.57 (d, J=16.6 Hz, 1H), 5.43 (br s, 1H), 3.32 (dt, J=7.0, 6.1 Hz, 2H),2.70 (t, J=8.0 Hz, 2H), 2.36 (s, 6H), 1.96 (s, 3H), 1.86-1.96 (m, 2H).

Example 52 Preparation of(E/Z)—N-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)pentadecanamide

(E/Z)—N-(3-(3-(2,6-dimethylstyryl)phenyl)propyl)pentadecanamide wasprepared according to the method used in Example 51.

Step 1:

To a solution of (E/Z)-3-(3-(2,6-dimethylstyryl)phenyl)propan-1-amine(isomer ratio 80:20 trans:cis, Example 1) (0.035 g, 0.132 mmol) inCH₂Cl₂ (2 mL) was added triethylamine (0.045 mL, 0.32 mmol), palmitoylchloride (0.045 mL, 0.15 mmol) and N, N-dimethylaminopyridine (˜2 mg).The reaction mixture was stirred at room temperature for 2 h. Additionaltriethylamine (0.045 mL, 0.32 mmol) and palmitoyl chloride (0.045 mL,0.15 mmol) were added and the mixture was stirred for 1 h. The mixturewas concentrated under reduced pressure and the residue was partitionedbetween EtOAc and water. The combined organics were washed with 5%aqueous NaHCO₃ and brine, dried over Na₂SO₄ and concentrated underreduced pressure. Purification by flash chromatography (0 to 75%EtOAc-hexanes gradient) gave Example 52 as a white semi-solid. Yield(0.060 g, 91%), trans-/cis-isomer ratio 5:1. Trans-isomer: ¹H NMR (400MHz, CDCl₃) δ 7.36 (d, J=7.6 Hz, 1H), 7.27-7.31 (m, 2H), 7.03-7.13 (m,3H), 6.58 (d, J=16.4 Hz, 1H), 3.33 (dt, J=7.0, 6.1 Hz, 2H), 2.69 (t,J=7.6 Hz, 2H), 2.36 (s, 6H), 2.12-2.16 (m, 2H), 1.86-1.90 (m, 2H),1.58-1.65 (m, 2H), 1.26 (m, 27H), 0.89 (t, J=6.8 Hz, 3H).

Example 53 Preparation of(E)-3-(2-methyl-5-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine

(E)-3-(2-methyl-5-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine(80:20 trans:cis) was prepared according to the method used in Example1.

Step 1:

To a −78° C. solution oftriphenyl((2,6,6-trimethylcyclohex-1-enyl)methyl)phosphonium bromide(24) (0.3787 g, 0.79 mmol) and 18-crown-6 (0.0246 g, 0.093 mmol) inCH₂Cl₂ (7 mL) was added potassium tert-butoxide (0.0991 g, 0.88 mmol)The mixture was sonicated at room temperature under argon for 2 min,resulting in a deep red solution. The mixture was cooled to −78° C. thena solution of 3-iodo-4-methylbenzaldehyde (0.1742 g, 0.71 mmol) inCH₂Cl₂ (3 mL+2 mL) was added and stirred for 5 min. The reaction mixturewas allowed to warm to room temperature. Additional phosphonium salt 24(0.3725 g, 1.6 mmol), 3-iodo-4-methylbenzaldehyde (0.2538 g, 1.7 mmol)and potassium tert-butoxide (0.1548 g, 1.38 mmol) were then added andthe reaction was stirred at room temperature for 4.3 h. The reaction waspartitioned between EtOAc and water then the combined organics werewashed with brine, dried over MgSO₄ and concentrated under reducedpressure. Purification by flash chromatography (6 to 50% EtOAc-hexanesgradient) gave(E)-2-iodo-1-methyl-4-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)benzeneas a white solid. Yield (0.26 g, 45%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.91(d, J=1.6 Hz, 1H), 7.44 (dd, J=7.6, 1.2 Hz, 1H), 7.29 (d, J=7.2 Hz, 1H),6.71 (d, J=16.4 Hz, 1H), 6.28 (6, J=16.8 Hz, 1H), 2.35 (s, 3H), 2.01 (t,J=6.4 Hz, 2H), 1.72 (s, 3H), 1.57-1.64 (m, 2H), 1.43-1.47 (m, 2H), 1.04(s, 6H).

Step 2:

(E)-2-iodo-1-methyl-4-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)benzenewas coupled with allyl alcohol following the method used in Example 32except that the reaction was stirred at room temperature overnight afterheating for 3 h at 60° C. Purification twice by flash chromatography (6to 25% EtOAc-hexanes gradient) gave(E/Z)-3-(2-methyl-5-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propanalas an oil. Yield (0.15 g, 71%); trans-/cis-isomer ratio 3:1.Trans-isomer: ¹H NMR (400 MHz, DMSO-d₆) δ 9.75 (t, J=1.2 Hz, 1H),7.19-7.27 (m, 2H), 7.09 (d, J=7.6 Hz, 1H), 6.64 (6, J=16.4 Hz, 1H), 6.27(d, J=16.8 Hz, 1H), 2.83 (t, J=5.4 Hz, 2H), 2.75 (t, J=5.4 Hz, 2H), 2.25(s, 3H), 2.01 (t, J=5.4 Hz, 2H), 1.74 (s, 3H), 1.58-1.63 (m, 2H),1.39-1.47 (m, 2H), 1.06 (s, 6H).

Step 3:

Reductive amination of(E/Z)-3-(2-methyl-5-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propanalwas conducted according to the method used in Example 46 except thatisopropanol-MeOH (1:2) was used as the solvent and the reaction wasstirred briefly at 0° C. after the addition of NaBH₄. Purification byflash chromatography (1:1 EtOAc/hexanes then 1:5:5 7 M NH₃ inMeOH/EtOAc/hexanes) gave Example 53 as an oil. Yield (0.1450 g, 96%): ¹HNMR (400 MHz, DMSO-d₆) δ 7.86 (br s, 3H), 7.21-7.23 (m, 2H), 7.11 (d,J=8.0 Hz, 1H), 6.64 (6, J=16.4 Hz, 1H), 6.27 (d, J=16.0 Hz, 1H),2.83-2.88 (m, 2H), 2.62 (t, J=8.0 Hz, 2H), 2.25 (s, 3H), 1.99-2.09 (m,2H), 1.76-1.84 (m, 2H), 1.72 (s, 3H), 1.57-1.61 (m, 2H), 1.45-1.47 (m,2H), 1.04 (s, 6H).

Example 54 Preparation of(E/Z)-4-amino-2-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-2-ol

(E/Z)-4-Amino-2-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-2-olwas prepared according to Scheme 20.

Step 1:

To a −78° C. solution of n-butyl lithium (2 mL of a 1.6 M solution inhexanes, 3.2 mmol) in THF (2 mL) was added a solution of iodide 31(0.4394 g, 1.35 mmol) in THF (5 mL+1 mL) and the reaction mixture wasstirred for 40 min. A solution of4-(tert-butyldimethylsilyloxy)butan-2-one (0.5040 g, 2.49 mmol) in THF(5 mL) was added slowly and the mixture was stirred for 35 min.Additional 4-(tert-butyldimethylsilyloxy)butan-2-one (0.1458 g, 0.72mmol) in THF (2 mL) was added. The mixture was stirred at −78° C. for 25min then warmed to room temperature. The reaction was quenched with theaddition of brine and extracted with EtOAc. The combined organics werewashed with brine, dried over MgSO₄ and concentrated under reducedpressure. Purification by flash chromatography (2 to 20% EtOAc-hexanesgradient) provided alcohol 59 as an oil. Yield (0.1870 g, 32%): ¹H NMR(400 MHz, DMSO-d₆) δ 7.56 (s, 1H), 7.29-7.38 (m, 3H), 6.76 (dd, J=16.4,0.8 Hz, 1H), 6.41 (d, J=16.4 Hz, 1H), 5.00 (s, 1H), 3.66-3.72 (m, 1H),3.44-3.50 (m, 1H), 2.09 (t, J=6.0 Hz, 2H), 2.03 (t, J=6.0 Hz, 2H), 1.80(s, 3H), 1.65-1.67 (m, 2H), 1.52-1.55 (m, 2H), 1.50 (s, 3H), 1.19 (s,6H), 0.87 (s, 9H), 0.00 (s, 6H).

Step 2:

To a solution of alcohol 59 (0.1870 g, 0.44 mmol) in THF (10 mL) wasadded a solution of tetrabutylammonium fluoride (1 mL of a 1 M solutionin THF, 1 mmol) The reaction mixture was stirred for 30 min thenconcentrated under reduced pressure. The residue was partitioned betweenEtOAc and water and the combined organics were washed with water andbrine. The solution was dried over MgSO₄ and concentrated under reducedpressure to give diol 60 as an oil. This material was taken on to thenext step without further purification. ¹H NMR (400 MHz, DMSO-d₆) δ 7.53(s, 1H), 7.26-7.37 (m, 3H), 6.75 (dd, J=16.4, 0.8 Hz, 1H), 6.39 (d,J=16.4 Hz, 1H), 5.05 (s, 1H), 4.37 (t, J=5.0 Hz, 1H), 3.42-3.51 (m, 1H),3.26-3.37 (m, 1H), 2.05 (t, J=7.2 Hz, 2H), 1.87-1.97 (m, 2H), 1.78 (s,3H), 1.63-1.66 (m, 2H), 1.49-1.52 (m, 2H), 1.46 (s, 3H), 1.20 (s, 6H).

Step 3:

To a solution of diol 60 (˜0.436 mmol) in CH₂Cl₂ (5 mL) was added N,N-dimethylaminopyridine (0.1362 g, 1.11 mmol) and a solution ofp-toluenesulfonyl chloride (0.0863 g, 0.75 mmol) in CH₂Cl₂ (3 mL). Thereaction mixture was stirred at room temperature for 25 min thenadditional p-toluenesulfonyl chloride (0.0288 g, 0.15 mmol) in THF (1mL) was added. The reaction was stirred for 1 h, 45 min and the mixturewas partitioned between EtOAc and saturated aqueous NaHCO₃. The combinedorganics were washed with more saturated aqueous NaHCO₃ and brine, driedover MgSO₄ and concentrated under reduced pressure to give tosylate 61as an oil. This material was taken on to the next synthetic step withoutfurther purification. Yield (0.2141 g crude, quant. for two steps): ¹HNMR (400 MHz, DMSO-d₆) δ 7.70-7.73 (m, 2H), 7.46-7.48 (m, 3H), 7.36 (d,J=7.2 Hz, 1H), 7.22-7.29 (m, 2H), 6.74 (d, J=16.4 Hz, 1H), 6.37 (d,J=16.4 Hz, 1H), 5.23 (s, 1H), 4.05-4.12 (m, 1H), 3.78-3.86 (m, 1H), 2.45(s, 3H), 2.11-2.32 (m, 4H), 1.78 (s, 3H), 1.62-1.68 (m, 2H), 1.48-1.56(m, 2H), 1.43 (s, 3H), 1.09 (s, 6H).

Step 4:

To a solution of tosylate 61 (˜0.4362 mmol) in DMF (5 mL) was addedpotassium phthalimide (0.1570 g, 0.85 mmol). The reaction mixture wasstirred at room temperature for 40 min then heated at 60° C. for 1 h.NaI (0.0831 g, 0.55 mmol) was added and heating was continued overnight.After cooling to room temperature, the mixture was concentrated underreduced pressure and partitioned between EtOAc and water. The combinedorganics were washed with water and brine, dried over MgSO₄ andconcentrated under reduced pressure. Purification by flashchromatography (20 to 50% EtOAc-hexanes gradient) gave phthalimide 62 asan oil. Yield (0.1179 g, 61% from alcohol 59): ¹H NMR (400 MHz, DMSO-d₆)δ 7.73-7.77 (m, 4H), 7.48 (s, 1H), 7.14-7.25 (m, 3H), 6.58 (d, J=16.0Hz, 1H), 6.29 (d, J=16.0 Hz, 1H), 5.12 (s, 1H), 3.56-3.61 (m, 1H),3.39-3.47 (m, 1H), 2.12-2.18 (m, 1H), 1.94-2.03 (m, 3H), 1.73 (s, 3H),1.56-1.62 (m, 2H), 1.42-1.56 (m, 5H), 1.05 (s, 6H).

Step 5:

Phthalimide 62 was deprotected according to the method used in Example32 to give Example 54 as an oil. Yield (0.0628 g, 81%): ¹H NMR (400 MHz,DMSO-d₆) δ 7.47 (s, 1H), 7.21-7.31 (m, 3H), 6.68 (dd, J=16.4, 0.8 Hz,1H), 6.34 (d, J=16.4 Hz, 1H), 2.40-2.60 (m, 4H), 2.00 (t, J=6.0 Hz, 2H),1.75-1.82 (m, 3H), 1.72 (s, 3H), 1.55-1.62 (m, 2H), 1.42-1.48 (m, 2H),1.37 (s, 3H), 1.03 (s, 6H).

Example 55 Preparation of(E)-3-fluoro-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine

(E)-3-Fluoro-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-aminewas prepared according to Scheme 21.

Step 1:

To a solution of (E)-3-amino-1-(3-(2,6-dimethylstyryl)phenyl)propan-1-ol(˜0.68 mmol) in THF (3 mL) was added a solution of di-tert-butyldicarbonate (0.1982 g, 0.91 mmol) in THF (3 mL). The reaction mixturewas stirred at room temperature for 10 min then the mixture wasconcentrated under reduced pressure to give crude alcohol 63. Thismaterial was used in the next synthetic step without furtherpurification. Yield (0.3137 g, quant.): ¹H NMR (400 MHz, CDCl₃) δ 7.39(s, 1H), 7.29-7.31 (m, 2H), 7.20 (dt, J=6.4, 2.0 Hz, 1H), 6.68 (dd,J=16.4, 0.8 Hz, 1H), 6.33 (d, J=16.4 Hz, 1H), 4.89 (s, 1H), 4.72-4.78(m, 1H), 3.43-3.58 (m, 1H), 3.12-3.22 (m, 2H), 2.02-2.05 (m, 2H),1.85-1.92 (m, 2H), 1.75 (s, 3H), 1.61-1.68 (m, 2H), 1.44-1.54 (m, 11H),1.06 (s, 6H).

Step 2:

To a −78° C. solution of crude alcohol 63 (˜0.6841 mmol) in CH₂Cl₂ (5mL) was added (diethylamino)sulfur trifluoride (0.15 mL, 1.14 mmol) Thereaction was stirred at −78° C. for 10 min then poured into a mixture ofEtOAc and saturated aqueous NaHCO₃. The layers were separated and theaqueous layer was extracted with EtOAc. The combined organics werewashed with brine, dried over MgSO₄ and concentrated under reducedpressure. Purification by flash chromatography (10 to 40% EtOAc-hexanesgradient) gave fluoride 64 as an oil. Yield (0.0946 g, 34%): ¹H NMR (400MHz, CDCl₃) δ 7.30-7.38 (m, 3H), 7.16 (d, J=7.6 Hz, 1H), 6.70 (d, J=16.0Hz, 1H), 6.34 (d, J=16.0 Hz, 1H), 5.53 (ddd, J=48.4, 8.8, 4.0 Hz, 1H),4.73 (br s, 1H), 3.24-3.40 (m, 2H), 2.08-2.20 (m, 2H), 2.04 (t, J=6.0Hz, 2H), 1.76 (s, 3H), 1.60-1.68 (m, 2H), 1.42-1.54 (m, 11H), 1.06 (s,6H).

Step 5:

Fluoride 64 was deprotected according to the method used in Example 45except that the reaction was stirred at room temperature overnight.Purification by flash chromatography (5:5:1 EtOAc:hexanes:7 M NH₃ inMeOH) gave Example 55 as an oil. Yield (0.0360 g, 51%): ¹H NMR (400 MHz,CD₃OD) δ 7.20-7.37 (m, 4H), 6.72 (d, J=16.0 Hz, 1H), 6.33 (d, J=16.0 Hz,1H), 5.56 (ddd, J=48.4, 8.8, 4.0 Hz, 1H), 2.76-2.82 (m, 2H), 1.91-2.20(m, 4H), 1.75 (s, 3H), 1.63-1.69 (m, 2H), 1.49-1.52 (m, 2H), 1.06 (s,6H).

Example 56 Preparation of(E)-3-amino-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-one

(E)-3-Amino-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-onewas prepared according to Scheme 22.

Step 1:

To a solution of alcohol 63 (0.3753 g, 0.94 mmol) in CH₂Cl₂ (15 mL) wasadded pyridinium chlorochromate (0.2662 g, 1.2 mmol) and Celite (0.45g). The reaction mixture was stirred for 1 h then the solids wereremoved by filtration through Celite. The filtrate was concentratedunder reduced pressure. Purification by flash chromatography (10 to 30%EtOAc-hexanes gradient) provided ketone 65 as an oil. Yield (0.1038 g,28%): ¹H NMR (400 MHz, CDCl₃) δ 7.95 (s, 1H), 7.77 (d, J=7.6 Hz, 1H),7.60 (d, J=7.6 Hz, 1H), 7.40 (t, J=7.6 Hz, 1H), 6.75 (dd, J=16.4, 0.8Hz, 1H), 6.37 (dd, J=16.4, 0.8 Hz, 1H), 5.16 (br s, 1H), 3.55 (q, J=6.0Hz, 2H), 3.21 (t, J=5.6 Hz, 2H), 2.04 (t, J=6.2 Hz, 2H), 1.75 (d, J=0.4Hz, 3H), 1.61-1.67 (m, 2H), 1.48-1.51 (m, 2H), 1.42 (s, 9H), 1.06 (s,6H).

Step 2:

To a solution of ketone 65 (0.1038 g, 0.26 mmol) in EtOAc (5 mL) wasadded a solution of HCl (5 mL of a ˜10 M solution in diethyl ether, 50mmol) and the mixture was stirred for 5 min. Additional HCl (10 mL of a˜10 M solution in diethyl ether, 100 mmol) was added and stirringcontinued for 5 min. The mixture was concentrated under reduced pressurethen co-evaporated under reduced pressure with EtOAc, EtOH, EtOAc,toluene, EtOAc-hexanes and EtOAc successively. Example 56 hydrochloridewas isolated as an oil. Yield (0.0974 g, quant.): ¹H NMR (400 MHz,CDCl₃) δ 8.35 (br s, 3H), 7.83 (s, 1H), 7.67 (d, J=7.2 Hz, 1H), 7.49 (d,J=7.6 Hz, 1H), 7.24 (t, J=7.2 Hz, 1H), 6.64 (d, J=16.0 Hz, 1H), 6.26 (d,J=16.4 Hz, 1H), 3.52 (s, 2H), 3.45 (s, 2H), 1.95 (t, J=6.0 Hz, 2H), 1.65(s, 3H), 1.52-1.58 (m, 2H), 1.39- 1.41 (m, 2H), 0.96 (s, 6H).

Example 57 Preparation of(E)-4-amino-1,1,1-trifluoro-2-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-2-ol

(E)-4-Amino-1,1,1-trifluoro-2-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-2-olwas prepared according to Scheme 23.

Step 1:

Phosphonium bromide 24 was coupled with 3-bromobenzaldehyde followingthe method used in Example 46. After stirring overnight, the reactionmixture was concentrated under reduced pressure. The residue wastriturated with ˜5% EtOAc-hexanes and the solids were removed byfiltration. The filtrate was concentrated under reduced pressure.Purification by flash chromatography (5% EtOAc-heptane) gave arylbromide 66 as an oil. Yield (3.52 g, 87%): ¹H NMR (400 MHz, CDCl₃) δ7.47 (t, J=2.0 Hz, 1H), 7.22-7.27 (m, 2H), 7.10 (t, J=7.6 Hz, 1H), 6.60(dd, J=16.0, 0.8 Hz, 1H), 6.19 (d, J=16.0 Hz, 1H), 1.97 (t, J=6.0 Hz,2H), 1.67 (s, 3H), 1.54-1.60 (m, 2H), 1.41-1.44 (m, 2H), 0.99 (s, 6H).

Step 2:

To a −78° C. solution of aryl bromide 66 in THF (7 mL) under argon wasadded a solution of n-butyl lithium (0.75 mL of a 1.6 M solution inhexane, 1.2 mmol) and the mixture was stirred for 20 min. Ethyltrifluoroacetate (0.215 mL, 1.8 mmol) was added and the reaction mixturewas allowed to warm to room temperature. The reaction was quenched withwater. The mixture was partitioned between EtOAc and water and thecombined organics were washed with brine, dried over MgSO₄ thenconcentrated under reduced pressure. Purification by flashchromatography (6 to 70% EtOAc-hexanes gradient) to give ketone 67 as alight yellow oil. Yield (0.200 g, 66%): ¹H NMR (400 MHz, DMSO-d₆) δ8.01-8.03 (m, 2H), 7.87 (dd, J=8.0, 1.2 Hz, 1H), 7.62 (t, J=8.0 Hz, 1H),6.86 (dd, J=16.4, 0.8 Hz, 1H), 6.51 (d, J=16.4 Hz, 1H), 2.02 (t, J=6.0,2H), 1.73 (s, 3H), 1.56-1.62 (m, 2H), 1.44-1.47 (m, 2H), 1.04 (s, 6H).

Step 3:

Reaction of ketone 67 with acetonitrile was conducted following themethod used in Example 50 except that lithium diisopropylamide was addedto the acetonitrile. Purification by flash chromatography (7 to 60%EtOAc-hexanes gradient) gave nitrile 68 as a colorless oil whichsolidified on standing. Yield (0.192 g, 80%): ¹H NMR (400 MHz, DMSO-d₆)δ 7.69 (s, 1H), 7.55 (d, J=8.0 Hz, 1H), 7.47-7.50 (m, 2H), 7.39 (t,J=7.6 Hz, 1H), 6.78 (d, J=15.6 Hz, 1H), 6.38 (d, J=16.4 Hz, 1H), 3.75(d, J=16.8 Hz, 1H), 3.37 (d, J=17.2 Hz, 1H), 2.02 (t, J=6.0 Hz, 1H),1.73 (s, 3H), 1.56-1.62 (m, 2H), 1.44-1.47 (m, 2H), 1.04 (s, 6H).

Step 4:

Nitrile 68 was reduced with LiAlH₄ following the method used in Example50 except that the reaction was stirred at 0° C. for 50 min. After thereaction was quenched, dried and concentrated under reduced pressure,the residue was suspended in hexanes and sonicated. The resultingsolution was stored at −20° C. for ˜3 h and sonicated once during thistime. The white crystals were collected by filtration and washed withhexanes to give Example 57 as a white solid (0.0495 g, 31% yield).Additional product was obtained by concentrating the mother liquor underreduced pressure to give a yellow oil (0.0604 g, 38% yield): ¹H NMR (400MHz, CDCl₃) δ 7.61 (s, 1H), 7.49 (d, J=7.6 Hz, 1H), 7.40 (d, J=7.6 Hz,1H), 7.34 (t, J=7.6 Hz, 1H), 6.72 (d, J=16.4 Hz, 1H), 6.37 (d, J=16.8Hz, 1H), 2.71-2.76 (m, 1H), 2.06-2.20 (m, 2H), 1.99-2.02 (m, 2H), 1.97(s, 3H), 1.61-1.67 (m, 2H), 1.48-1.52 (m, 2H), 1.07 (s, 6H). Onealiphatic proton is obscured by the peak for residual protonated DMSO.

Example 58 Preparation of(E)-3-amino-2,2-dimethyl-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-ol

(E)-3-Amino-2,2-dimethyl-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-olwas prepared according to Scheme 24.

Step 1:

To a −78° C. solution of n-butyl lithium (2.5 mL of a 1.6 M solution inhexanes, 4.0 mmol) in THF (8 mL) was added a solution of aryl bromide 66(0.9152 g, 3.0 mmol) in THF (12 mL). The reaction was stirred at −78° C.for 10 min then DMF (0.6 mL, 7.7 mmol) was added and the reaction wasstirred for 5 min then allowed to warm to ˜−60° C. The reaction wasstirred for 5 min then quenched with saturated aqueous NH₄Cl. Afterwarming to room temperature, the layers were separated and the aqueouslayer was extracted with EtOAc. The combined organics were washed withbrine, dried over MgSO₄ and concentrated under reduced pressure.Purification by flash chromatography (5 to 20% EtOAc-hexanes gradient)gave aldehyde 69 as an oil. Yield (0.6467 g, 85%): ¹H NMR (400 MHz,CDCl₃) δ 10.0 (s, 1H), 7.90 (t, J=1.6 Hz, 1H), 7.72 (dt, J=7.6, 1.2 Hz,1H), 7.65 (d, J=7.8 Hz, 1H), 7.48 (t, J=7.6 Hz, 1H), 6.79 (dd, J=16.2,1.0 Hz, 1H), 6.40 (d, J=16.4 Hz, 1H), 2.04-2.07 (m, 2H), 1.77 (s, 3H),1.62-1.68 (m, 2H), 1.49-1.52 (m, 2H), 1.07 (s, 6H).

Step 2:

Aldehyde 69 was reacted with isobutyronitrile according to the methodused in Example 50. Purification by flash chromatography (10 to 70%EtOAc-hexanes gradient) gave nitrile 70 as an oil. (Yield (0.2235 g,69%): ¹H NMR (400 MHz, CDCl₃) δ 7.26-7.43 (m, 4H), 6.72 (dd, J=16.0, 0.8Hz, 1H), 6.35 (d, J=16.0 Hz, 1H), 4.55 (d, J=3.2 Hz, 1H), 2.05-2.08 (m,2H), 1.76 (s, 3H), 1.61-1.67 (m, 2H), 1.48-1.51 (m, 2H), 1.45 (s, 3H),1.25 (s, 3H), 1.07 (s, 6H).

Step 3:

Nitrile 70 was reduced with LiAlH₄ according to the method used inExample 50 except that the reaction was stirred at 0° C. for 1.25 h.Purification by flash chromatography (2:9:9 (7 M NH₃ inMeOH)/EtOAc/hexanes) gave Example 58 as an oil. Yield (0.1750 g, 77%):¹H NMR (400 MHz, CD₃OD) δ 7.23-7.36 (m, 3H), 7.17 (dd, J=7.6, 1.2 Hz,1H), 6.70 (dd, J=16.0, 0.8 Hz, 1H), 6.33 (d, J=16.0 Hz, 1H), 4.55 (s,1H), 2.73 (d, J=13.2 Hz, 1H), 2.55 (d, J=13.2 Hz, 1H), 2.05 (t, J=6.4Hz, 2H), 1.75 (s, 3H), 1.60-1.71 (m, 2H), 1.50-1.53 (m, 2H), 1.06 (s,6H), 0.84 (s, 6H).

Example 59 Preparation of(E)-(syn/anti)-3-amino-2-methyl-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-ol

(E)-3-Amino-2-methyl-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-olwas prepared according to the method used in Example 58.

Step 1:

Aldehyde 69 was reacted with propionitrile according to the method usedin Example 58. Purification twice by flash chromatography (10 to 70%EtOAc-hexanes gradient; then 10 to 30% EtOAc-hexanes gradient) gave bothdiastereomers of(E)-3-hydroxy-2-methyl-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propanenitrile.The first eluting diastereomer was isolated as an oil (0.0233 g). Thesecond eluting diastereomer was isolated as an oil (0.1214 g). Totalyield (0.1447 g, 47%):

Diastereomer 2: ¹H NMR (400 MHz, CDCl₃) δ 7.32-7.41 (m, 3H), 7.20-7.26(m, 1H), 6.72 (d, J=16.4 Hz, 1H), 6.35 (dd, J=16.4, 1.2 Hz, 1H), 4.81(dd, J=6.0, 3.2 Hz, 1H), 3.01 (quint, J=7.2 Hz, 1H), 2.59 (d, J=3.2 Hz,1H), 2.04 (t, J=6.4 Hz, 2H), 1.76 (s, 3H), 1.65 (quint, J=6.4 Hz, 2H),1.50 (t, J=6.0 Hz, 2H), 1.28 (d, J=7.2 Hz, 3H), 1.07 (s, 6H).

Step 2:

Diastereomer 2 of(E)-3-hydroxy-2-methyl-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propanenitrilewas reduced with LiAlH₄ according to the method used in Example 58.Purification by flash chromatography (7 M NH₃ in MeOH/EtOAc/hexanes2:9:9) gave both diastereomers of Example 59 as an oil (2.8:1diastereomeric mixture). Yield (0.0893 g, 73%). Major diastereomer: ¹HNMR (400 MHz, CDCl₃) δ 7.26-7.38 (m, 3H), 7.18 (dt, J=6.8, 1.6 Hz, 1H),6.68 (dd, J=16.0, 0.8 Hz, 1H), 6.35 (d, J=16.0 Hz, 1H), 4.98 (d, J=3.2Hz, 1H), 2.94-2.96 (m, 2H), 2.03 (t, J=6.4 Hz, 2H), 1.98-2.01 (m, 1H),1.76 (s, 3H), 1.64 (quint, J=6.4 Hz, 2H), 1.49 (t, J=6.0 Hz, 2H), 1.06(s, 6H), 0.84 (d, J=7.2 Hz, 3H).

Example 60 Preparation of(E)-3-amino-2,2-dimethyl-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-one

(E)-3-Amino-2,2-dimethyl-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-onewas prepared according to the methods used in Examples 55 and 56.

Step 1:

(E)-3-Amino-2,2-dimethyl-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-ol(Example 58) was protected with di-tert-butyl dicarbonate according tothe method used in Example 55 to give (E)-tert-butyl3-hydroxy-2,2-dimethyl-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propylcarbamateas an oil. This compound was used in the next synthetic step withoutpurification. Yield (0.1495 g, quant.).

Step 2:

(E)-Tert-butyl3-hydroxy-2,2-dimethyl-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propylcarbamate(0.0809 g, 0.19 mmol) was reacted with pyridinium chlorochromateaccording to the method used in Example 56 except that the reaction timewas 2 h, 15 min. Purification by flash chromatography (10 to 40%EtOAc-hexanes gradient) gave (E)-tert-butyl2,2-dimethyl-3-oxo-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propylcarbamateas an oil. ¹H NMR (400 MHz, CD₃OD) δ 7.72 (s, 1H), 7.58 (d, J=7.6 Hz,1H), 7.52 (d, J=8.0 Hz, 1H), 7.35 (t, J=7.6 Hz, 1H), 6.72 (dd, J=16.0,0.8 Hz, 1H), 6.33 (d, J=16.0 Hz, 1H), 5.05 (t, J=6.0 Hz, 1H), 3.38 (d,J=6.8 Hz, 2H), 2.04 (t, J=6.4 Hz, 2H), 1.76 (s, 3H), 1.61-1.67 (m, 2H),1.48-1.50 (m, 2H), 1.36-1.42 (m, 15H), 1.06 (s, 6H).

Step 3:

To a solution of (E)-tert-butyl2,2-dimethyl-3-oxo-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propylcarbamate(˜0.19 mmol) in EtOAc (2 mL) was added a solution of HCl (10 mL of a ˜10M solution in diethyl ether, 100 mmol) and the mixture was stirred for10 min. Additional HCl (5 mL of a ˜10 M solution in diethyl ether, 50mmol) was added and the reaction was stirred for 10 min. The mixture wasconcentrated under reduced pressure to give Example 60 hydrochloride asan oil. Yield (0.0500 g, 73% for two steps): ¹H NMR (400 MHz, CD₃OD) δ9.52 (br s, 3H), 7.72 (s, 1H), 7.58 (d, J=7.6 Hz, 1H), 7.52 (d, J=8.0Hz, 1H), 7.31 (t, J=7.6 Hz, 1H), 6.71 (d, J=16.0 Hz, 1H), 6.32 (d,J=16.0 Hz, 1H), 3.28 (br s, 2H), 2.02 (t, J=6.4 Hz, 2H), 1.73 (s, 3H),1.46-1.66 (m, 10H), 1.04 (s, 6H).

Example 61 Preparation of(E)-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propane-1,3-diamine

(E)-1-(3-(2-(2,6,6-Trimethylcyclohex-1-enyl)vinyl)phenyl)propane-1,3-diaminewas prepared according to Scheme 25.

Step 1:

To a solution of alcohol 63 (0.2473 g, 0.62 mmol) in THF (7 mL) wasadded triphenylphosphine (0.3540 g, 1.35 mmol), phthalimide (0.2080 g,1.41 mmol) and a solution of diethyl azodicarboxylate (0.2905 g, 1.67mmol) in THF (3 mL). The reaction was stirred at room temperature for 5min, then at 60° C. for 1 h. Additional triphenylphosphine (0.1369 g,0.52 mmol), phthalimide (0.0974 g, 0.66 mmol) and diethylazodicarboxylate (100.4, 0.64 mmol) were added and the mixture wasstirred at 50° C. overnight. The mixture was concentrated under reducedpressure. Purification by flash chromatography (10 to 70% EtOAc-hexanesgradient) provided phthalimide 71 as an oil. Yield 0.1659 g, 50%): ¹HNMR (400 MHz, CDCl₃) δ 7.64-7.84 (m, 4H), 7.48 (s, 1H), 7.41 (d, J=7.6Hz, 1H), 7.34 (d, J=8.0 Hz, 1H), 7.24-7.29 (m, 1H), 6.67 (dd, J=16.0,0.8 Hz, 1H), 6.30 (d, J=16.0 Hz, 1H), 5.41 (dd, J=10.0, 5.6 Hz, 1H),4.72 (br s, 1H), 3.22-3.32 (m, 1H), 3.08-3.17 (m, 1H), 2.76-2.88 (m,1H), 2.40-2.48 (m, 1H), 2.01 (t, J=6.0 Hz, 2H), 1.73 (s, 3H), 1.59-1.65(m, 2H), 1.46-1.48 (m, 2H), 1.39 (s, 9H), 1.03 (s, 6H).

Step 2:

Phthalimide 71 was deprotected with hydrazine according to the methodused in Example 32 except that the reaction was heated to reflux for 2.5h. Purification by flash chromatography (7 M NH₃ in MeOH/EtOAc/hexanes2:9:9) provided amine 72 as an oil. Yield 0.1719 g, 68%): ¹H NMR (400MHz, CDCl₃) δ 7.26-7.31 (m, 3H), 7.13-7.17 (m, 1H), 6.68 (dd, J=16.0,0.8 Hz, 1H), 6.32 (d, J=16.0 Hz, 1H), 4.99 (br s, 1H), 3.96 (dt J=6.0,1.6 Hz, 1H), 3.15-3.27 (m, 2H), 2.04 (t, J=6.4 Hz, 2H), 1.78-1.89 (m,2H), 1.75 (s, 3H), 1.60-1.67 (m, 2H), 1.55 (br s, 2H), 1.41-1.51 (m,11H), 1.06 (s, 6H).

Step 3:

To a solution of amine 72 in EtOAc (2 mL) was added a solution of HCl(20 mL of a ˜10 M solution in diethyl ether, 200 mmol) The reactionmixture was stirred at room temperature for 30 min then the solid wascollected by filtration and washed with diethyl ether. The product wasdried under vacuum to give Example 61 dihydrochloride as a whitecrystalline solid. Yield (0.0574 g, 62%): ¹H NMR (400 MHz, DMSO-d₆) δ8.72 (br s, 3H), 8.03 (br s, 3H), 7.76 (s, 1H), 7.37-7.47 (m, 3H), 6.83(d, J=16.4 Hz, 1H), 6.35 (d, J=16.4 Hz, 1H), 4.39 (br s, 1H), 2.74-2.84(m, 1H), 2.26-2.37 (m, 2H), 2.12-2.24 (m, 1H), 2.02 (t, J=6.4 Hz, 2H),1.73 (s, 3H), 1.57-1.61 (m, 2H), 1.44-1.48 (m, 2H), 1.04 (s, 6H).

Example 62 Preparation of(E)-4,4,4-trifluoro-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-1-amine

(E)-4,4,4-Trifluoro-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-1-aminewas prepared according to Scheme 26.

Step 1:

To a −78° C. solution of 2-(3-bromophenyl)-1,3-dioxolane (73) (2.29 g,10.0 mmol) in THF (20 mL) under argon was added n-butyl lithium (7.5 mLof a 1.6 M solution in hexane, 12 mmol) The reaction mixture was stirredat −78° C. for 20 min then ethyl trifluoroacetate (2.2 mL, 18.0 mmol)was added rapidly. The reaction was allowed to warm to room temperatureover 45 min then quenched with water. The mixture was partitionedbetween EtOAc and water and the combined organics were washed withbrine, dried over Na₂SO₄ and concentrated under reduced pressure.Purification by flash chromatography (3 to 30% EtOAc-hexanes gradient)gave ketone 74 (contaminated with ˜10% 73) as an oil. (1.83 g, 80%): ¹HNMR (400 MHz, DMSO-d₆) δ 8.04-8.06 (m, 2H), 7.89 (dt, J=7.6, 1.2 Hz,1H), 7.70 (dd, J=8.4, 7.6 Hz, 1H), 5.87 (s, 1H), 4.04-4.08 (m, 2H),3.96-4.02 (m, 2H).

Step 2:

To an ice cold solution of triethyl phosphonoacetate (1.74 mL, 8.7 mmol)in THF under argon was added n-butyl lithium (6 mL of a 1.6 M solutionin hexane, 9.6 mmol) over a ˜5 min period. The reaction was allowed towarm to room temperature and stirred for 30 min. A solution of ketone 74(1.80 g, 7.9 mmol) in THF (3 mL) was added dropwise and the reactionmixture was stirred for 5 h. The mixture was partitioned between EtOAcand water and the combined organics were dried over Na₂SO₄ andconcentrated under reduced pressure. Purification by flashchromatography (7 to 60% EtOAc-hexanes gradient) gave alkene 75 (˜9:1isomer ratio) as a colorless oil. (Yield 1.62 g, 65%); trans-isomer: ¹HNMR (400 MHz, DMSO-d₆) δ 7.52 (dt, J=6.4, 1.2 Hz, 1H), 7.44-7.48 (m,1H), 7.29-7.31 (m, 2H), 6.89 (d, J=1.2 Hz, 1H), 5.74 (s, 1H), 4.02-4.05(m, 2H), 3.92-4.00 (m, 4H), 0.93 (t, J=7.2 Hz, 3H).

Step 3:

To a degassed solution of alkene 75 in EtOH (abs, 20 mL) under argon wasadded 10% Pd/C (˜20 mg). The reaction was placed under H₂ atmosphere andstirred for 5 h. The mixture was degassed, filtered through Celite, andthe filtrate was concentrated under reduced pressure. Ethyl3-(3-(1,3-dioxolan-2-yl)phenyl)-4,4,4-trifluorobutanoate was isolated asa colorless oil and used in the next synthetic step withoutpurification. Yield (0.92 g, 97%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.39-7.46(m, 4H), 5.71 (s, 1H), 3.94-4.04 (m, 7H), 2.95-3.09 (m, 2H), 1.03 (t,J=6.8 Hz, 3H).

Step 4:

To a solution of ethyl3-(3-(1,3-dioxolan-2-yl)phenyl)-4,4,4-trifluorobutanoate (0.92 g, 2.89mmol) in acetone (7 mL) and water (3 mL) was added p-toluenesulfonicacid (0.050 g, 0.26 mmol) The mixture was stirred at room temperaturefor 2.5 h then the volatiles were removed by concentration under reducedpressure. The residue was partitioned between EtOAc and water and thecombined organics were washed with saturated aqueous NaHCO₃, water andbrine. The solution was dried over a mixture of MgSO₄ and Na₂SO₄ andconcentrated under reduced pressure. Purification by flashchromatography (7 to 60% EtOAc-hexanes gradient) gave aldehyde 76 as acolorless oil. Yield (0.589 g, 74%): ¹H NMR (400 MHz, DMSO-d₆) δ 10.00(s, 1H), 7.97 (s, 1H), 7.89 (dt, J=7.6, 1.2 Hz, 1H), 7.79 (d, J=8.0 Hz,1H), 7.62 (t, J=7.6 Hz, 1H), 4.20-4.31 (m, 1H), 3.90-4.02 (m, 2H),3.09-3.11 (m, 2H), 1.03 (t, J=6.8 Hz, 3H).

Step 5:

Aldehyde 76 was coupled with phosphonium salt 24 according to the methodused in Example 46 except that the initial addition of potassiumtert-butoxide was conducted at −78° C. instead of 0° C. After stirringovernight, the reaction mixture was concentrated under reduced pressureand the residue was partitioned between EtOAc and water. The combinedorganics were washed with brine, dried over MgSO₄ and concentrated underreduced pressure. Purification by flash chromatography (7 to 60%EtOAc-hexanes gradient) gave alkene 77 as a colorless oil. Yield (0.833g, 98%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.51 (s, 1H), 7.45 (d, J=7.6 Hz,1H), 7.31 (t, J=7.6 Hz, 1H), 7.25 (d, J=7.6 Hz, 1H), 6.76 (d, J=16.8 Hz,1H), 6.34 (d, J=16.4 Hz, 1H), 3.93-4.09 (m, 3H), 2.98-3.10 (m, 2H), 2.01(t, J=6.4 Hz, 2H), 1.72 (s, 3H), 1.55- 1.61 (m, 2H), 1.43-1.46 (m, 2H),1.03-1.07 (m, 9H).

Step 6:

To an ice-cold solution of alkene 77 (0.522 g, 1.32 mmol) in THF (10 mL)under argon was added a solution of LiAlH₄ (0.75 mL of a 2.0 M solutionin THF, 1.5 mmol) The reaction was stirred at 0° C. for 30 min thendiluted with diethyl ether and quenched with saturated aqueous Na₂SO₄.The solution was dried over Na₂SO₄ and MgSO₄ and filtered throughCelite. The filtrate was concentrated under reduced pressure to give(E)-4,4,4-trifluoro-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-1-olas a colorless oil. This compound was used in the next synthetic stepwithout purification. Yield (0.470 g, quant.): ¹H NMR (400 MHz, DMSO-d₆)δ 7.48 (d, J=7.6 Hz, 1H), 7.41 (s, 1H), 7.34 (t, J=7.6 Hz, 1H), 7.21 (d,J=7.6 Hz, 1H), 6.75 (d, J=16.4 Hz, 1H), 6.35 (d, J=16.4 Hz, 1H), 4.61(dt, J=5.2, 0.4 Hz, 1H), 3.71 (ddd, J=9.6, 9.6, 4.4 Hz, 1H), 3.33-3.40(m, 1H), 3.10-3.18 (m, 1H), 2.00-2.07 (m, 4H), 1.73 (s, 3H), 1.56-1.62(m, 2H), 1.44-1.47 (m, 2H), 1.04 (s, 6H).

Step 7:

To an ice-cold solution of(E)-4,4,4-trifluoro-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-1-ol(0.470 g, 1.32 mmol) in THF (10 mL) was added phthalimide (0.200 g, 1.35mmol), triphenylphosphine (0.354 g, 1.35 mmol) and diethylazodicarboxylate (0.213 mL, 1.35 mmol) The mixture was allowed to warmto room temperature and stirred for 48 h. The mixture was concentratedunder reduced pressure and the residue was purified by flashchromatography (7 to 60% EtOAc-hexanes gradient) to give phthalimide 78as a colorless oil. Yield (0.479 g, 75%): ¹H NMR (400 MHz, DMSO-d₆) δ7.72-7.78 (m, 4H), 7.39 (s, 1H), 7.29 (d, J=7.6 Hz, 1H), 7.24 (t, J=7.6Hz, 1H), 7.17 (d, J=7.6 Hz, 1H), 6.71 (d, J=16.4 Hz, 1H), 6.28 (d,J=16.4 Hz, 1H), 3.72-3.81 (m, 1H), 3.46-3.59 (m, 2H), 2.26-2.33 (m, 2H),2.02 (t, J=6.0 Hz, 2H), 1.74 (s, 3H), 1.56-1.62 (m, 2H), 1.44-1.47 (m,2H), 1.06 (s, 6H).

Step 8:

To a solution of phthalimide 78 (0.475 g, 1.0 mmol) in EtOH (abs., 10mL) was added hydrazine hydrate (0.145 mL, 3.0 mmol) The reaction wasstirred at 70° C. overnight under argon. After cooling to roomtemperature, the reaction mixture was concentrated under reducedpressure. The residue was suspended in EtOAc, dried over MgSO₄,sonicated, and the solids were removed by filtration. The filtrate wasconcentrated under reduced pressure. A mixture of EtOAc-hexanes (50%)was added to the residue and the solids were removed by filtration. Thefiltrate was concentrated under reduced pressure to give Example 62 as acolorless oil. (Yield 0.322 g, 92%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.46(d, J=7.6 Hz, 1H), 7.42 (s, 1H), 7.32 (t, J=7.6 Hz, 1H), 7.20 (d, J=7.6Hz, 1H), 6.74 (d, J=16.4 Hz, 1H), 6.34 (d, J=16.4 Hz, 1H), 3.71-3.77 (m,1H), 2.42-2.45 (m, 1H), 2.24-2.31 (m, 1H), 2.01 (t, J=6.0 Hz, 2H),1.90-1.96 (m, 2H), 1.72 (s, 3H), 1.55-1.61 (m, 4H), 1.43-1.46 (m, 2H),1.04 (s, 6H).

Example 63 Preparation of(E)-3-methoxy-N-methyl-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine

(E)-3-Methoxy-N-methyl-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-aminewas prepared according to Scheme 27.

Step 1:

To a solution of alcohol 63 (0.2586 g, 0.65 mmol) in DMSO (3 mL) wasadded iodomethane (0.07 mL, 1.12 mmol) and powdered KOH (0.0989 g, 1.76mmol) The reaction was stirred at room temperature for 35 min thenpartitioned between EtOAc and water. The combined organics were washedwith brine, dried over MgSO₄ and concentrated under reduced pressure.Purification by flash chromatography (10 to 30% EtOAc-hexanes gradient)afforded methyl amine 79 (0.0511 g, 18% yield) and ether 80 (0.1074 g,40% yield) as oils.

Compound 79: ¹H NMR (400 MHz, CDCl₃) δ 7.28-7.34 (m, 3H), 7.13 (d, J=7.2Hz, 1H), 6.69 (d, J=16.4 Hz, 1H), 6.34 (d, J=16.4 Hz, 1H), 4.10 (dd,J=8.4, 4.8 Hz, 1H), 3.35 (br s, 2H), 3.23 (s, 3H), 2.85 (br s, 3H), 2.04(t, J=6.0 Hz, 2H), 1.85-1.98 (m, 2H), 1.76 (s, 3H), 1.61-1.67 (m, 2H),1.48-1.51 (m, 2H), 1.43 (s, 9H), 1.07 (s, 6H).

Compound 80: ¹H NMR (400 MHz, CDCl₃) δ 7.28-7.34 (m, 3H), 7.13 (dt,J=7.2, 1.6 Hz, 1H), 6.69 (dd, J=16.4, 0.8 Hz, 1H), 6.33 (d, J=16.4 Hz,1H), 4.19 (dd, J=8.4, 4.8 Hz, 1H), 3.18-3.22 (m, 5H), 2.04 (t, J=6.0 Hz,2H), 1.83-1.96 (m, 2H), 1.76 (s, 3H), 1.60-1.67 (m, 2H), 1.48-1.51 (m,2H), 1.44 (s, 9H), 1.07 (s, 6H).

Step 2:

Methyl amine 79 was deprotected according to the method used in Example45. Purification by flash chromatography (2:9:9 (7 M NH₃ inMeOH)/EtOAc/hexanes) afforded Example 63 as an oil in ˜80% purity. Yield(0.0349 g, 83%): ¹H NMR (400 MHz, CDCl₃) δ 7.26-7.34 (m, 3H), 7.14 (dt,J=7.2, 1.6 Hz, 1H), 6.70 (dd, J=16.4, 0.8 Hz, 1H), 6.33 (d, J=16.4 Hz,1H), 4.23 (dd, J=8.0, 5.2 Hz, 1H), 3.23 (s, 3H), 2.68 (t, J=6.8 Hz, 1H),2.43 (s, 3H), 1.96-2.05 (m, 4H), 1.79-1.89 (m, 1H), 1.76 (s, 3H),1.61-1.67 (m, 2H), 1.41-1.50 (m, 2H), 1.06 (s, 6H).

Example 64 Preparation of(E)-3-methoxy-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-amine

(E)-3-Methoxy-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-aminewas prepared according to Scheme 27. Step 1: Methyl amine 80 wasdeprotected according to the method used in Example 45. Purification byflash chromatography (7 M NH₃ in MeOH)/EtOAc/hexanes 2:9:9) affordedExample 64 as an oil in ˜80% purity. Yield (0.0708 g, quant.): ¹H NMR(400 MHz, CDCl₃) δ 7.25-7.34 (3H), 7.13-7.18 (m, 1H), 6.69 (dd, J=16.4,0.8 Hz, 1H), 6.33 (d, J=16.4 Hz, 1H), 4.24 (dd, J=8.4, 5.2 Hz, 1H), 3.23(s, 3H), 2.82 (t, J=6.8 Hz, 2H), 2.04 (t, J=8.4 Hz, 2H), 1.94-2.00 (m,1H), 1.85 (s, 3H), 1.77-1.83 (m, 1H), 1.61-1.67 (m, 2H), 1.47-1.50 (m,2H), 1.06 (s, 6H).

Example 65 Preparation of(E)-4-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)butan-2-amine

(E)-4-(3-(2-(2,6,6-Trimethylcyclohex-1-enyl)vinyl)phenyl)butan-2-aminewas prepared according to Scheme 28.

Step 1:

A solution of aryl bromide 73 (2.0 g, 8.8 mmol), 3-buten-2-one (0.926 g,13.2 mmol), NaHCO₃ (1.4 g, 13.2 mmol), PdCl₂ (PPh₃)₂ (0.309 g, 0.44mmol) and tri(o-tolyl)phosphine (0.134 g, 0.44 mmol) in DMF (10 mL) washeated to reflux under argon for 2 h. After cooling to room temperature,solids were removed by filtration. The filtrate was diluted with CH₂Cl₂and water. The combined organics were washed with water and brine, driedover MgSO₄ and concentrated under reduced pressure. Purification byflash chromatography (20 to 50% EtOAc-hexanes gradient) afforded alkene81 as a yellow oil. Yield (1.5 g, 79%): ¹H NMR (400 MHz, CDCl₃) δ7.67-7.69 (m, 1H), 7.50-7.55 (m, 3H), 7.43 (t, J=7.6 Hz, 1H), 6.74 (t,J=16.2 Hz, 1H), 5.82 (s, 1H), 4.01-4.16 (m, 4H), 2.38 (s, 3H).

Step 2:

To a solution of alkene 81 (1.5 g, 6.9 mmol) in MeOH (90 mL) was addedNiCl₂.H₂O (0.894 g, 6.9 mmol) then NaBH₄ (0.778 g, 20.6 mmol)portionwise. The mixture was stirred at room temperature for 15 min thenthe solids were removed by filtration. The filtrate was concentratedunder reduced pressure and the residue was partitioned between CH₂Cl₂and saturated aqueous NH₄Cl. The combined organics were dried over MgSO₄and concentrated under reduced pressure. Purification by flashchromatography (20 to 50% EtOAc-hexanes gradient) provided alcohol 82 asa colorless oil. Yield (0.911 g, 60%).

Step 3:

Alcohol 82 was coupled with phthalimide according to the general methodused in Example 32 to give2-(4-(3-(1,3-dioxolan-2-yl)phenyl)butan-2-yl)isoindoline-1,3-dione as awhite oily solid containing ˜10% phthalimide. Yield (0.686 g, 67%): ¹HNMR (400 MHz, CDCl₃) δ 7.79 (dd, J=6.6, 2.8 Hz, 2H), 7.69 (dd, J=6.6,2.8 Hz, 2H), 7.20-7.26 (m, 3H), 7.16-7.30 (m, 1H), 5.74 (s, 1H),4.38-4.45 (m, 1H), 4.01-4.11 (m, 4H), 2.43-2.68 (m, 3H), 2.00-2.06 (m,1H), 1.48 (d, J=6.8 Hz, 3H).

Step 4:

2-(4-(3-(1,3-Dioxolan-2-yl)phenyl)butan-2-yl)isoindoline-1,3-dione wasdeprotected according to the method used in Example 62 to give aldehyde83 as a colorless oil. This compound was used in the next synthetic stepwithout purification. Yield (0.544 g, quant.): ¹H NMR (400 MHz, DMSO-d₆)δ 9.88 (s, 1H), 7.76-7.81 (m, 4H), 7.64 (s, 1H), 7.58 (dt, J=7.2, 1.2Hz, 1H), 7.47 (dt, J=7.6, 1.2 Hz, 1H), 7.40 (t, J=7.6 Hz, 1H). 4.22-4.27(m, 1H), 2.57-2.70 (m, 2H), 2.31-2.41 (m, 1H), 1.99-2.06 (m, 1H), 1.39(d, J=7.2 Hz, 3H).

Step 5:

Aldehyde 83 was coupled with phosphonium salt 24 following the procedurein Example 44 except that the reaction mixture was stirred at roomtemperature for 3 h. Solids were removed by filtration through a pad ofsilica gel. The filtrate was concentrated under reduced pressure and theresidue was purified by flash chromatography (0 to 20% EtOAc-hexanesgradient) to give alkene 84 as an oil. Yield (0.568 g, 75%): ¹H NMR (400MHz, DMSO-d₆) δ 7.74-7.81 (m, 4H), 7.18 (br s, 1H), 7.10-7.11 (m, 2H),6.95-6.97 (m, 1H), 6.62 (dd, J=16.0, 0.8 Hz, 1H), 6.22 (d, J=16.0 Hz,1H), 4.21-4.27 (m, 1H), 2.49-2.59 (m, 1H), 2.33-2.48 (m, 2H), 1.94-2.01(m, 3H), 1.70 (s, 3H), 1.54-1.60 (m, 2H), 1.42-1.45 (m, 2H), 1.40 (d,J=5.6 Hz, 3H), 1.00 (s, 6H).

Step 6:

To a solution of alkene 84 (0.500 g, 1.2 mmol) in EtOH (5 mL) was addedhydrazine hydrate (0.30 g, 6.0 mmol) The reaction mixture was heated at50-60° C. for 2 h. After cooling to room temperature, the mixture wasconcentrated under reduced pressure. The residue was partitioned betweenCH₂Cl₂ and water and the combined organics were dried over MgSO₄ andconcentrated under reduced pressure. The crude material was purified byfiltration through a pad of silica gel (0-5% MeOH—CH₂Cl₂ then 5% 7 M NH₃in MeOH—CH₂Cl₂) to give Example 65 as a yellow oil. Yield (0.1970 g,56%), trans-/cis-isomer ratio 15.6:1. Trans-isomer: ¹H NMR (400 MHz,DMSO-d₆) δ 7.24-7.27 (m, 2H), 7.20 (t, J=8.0 Hz, 1H), 7.03 (d, J=7.2 Hz,1H), 6.68 (d, J=16.4 Hz, 1H), 6.29 (d, J=16.0 Hz, 1H), 2.76-2.77 (m,1H), 2.56-2.59 (m, 2H), 2.0 (t, J=6.1 Hz, 2H), 1.70 (s, 3H), 1.40-1.64(m, 6H), 0.99-1.02 (m, 9H).

Example 66 Preparation of(E)-1-amino-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-2-ol

(E)-1-Amino-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-2-olwas prepared according to Scheme 29.

Step 1:

To a −78° C. solution of aryl bromide 66 (0.50 g, 1.64 mmol) in THF (4mL) was added n-butyl lithium (1.1 mL of a 1.6 M solution in hexanes,1.76 mmol) dropwise. The mixture was stirred for 10 min then BF₃-diethyletherate (0.23 mL, 1.8 mmol) was added dropwise. After stirring for 3min, a solution of epichlorohydrin (0.11 mL, 1.4 mmol) in THF (1 mL) wasadded dropwise over 3 min. The reaction mixture was stirred for 45 minat −78° C. then quenched with the dropwise addition of water. Afterwarming to room temperature, the mixture was partitioned between MTBEand water. The combined organics were washed with water and brine, driedover Na₂SO₄ and concentrated under reduced pressure. Purification byflash chromatography (1:8 EtOAc: heptane) gave chlorohydrin 85. Yield(0.24 g, 57%): ¹H NMR (400 MHz, CDCl₃) δ 7.28-7.31 (m, 2H), 7.22 (t,J=8.0 Hz, 1H), 7.07 (d, J=8.0 Hz, 1H), 6.69 (dd, J=16.4, 0.8 Hz, 1H),6.30 (d, J=16.4 Hz, 1H), 5.16 (d, J=5.2 Hz, 1H), 3.86-3.89 (m, 1H), 3.56(dd, J=10.8, 4.8 Hz, 1H), 3.47 (dd, J=10.8, 5.6 Hz, 1H), 2.79 (dd,J=13.6, 5.2 Hz, 1H), 2.65 (dd, J=13.6, 5.2 Hz, 1H), 2.00 (t, J=6.0 Hz,2H), 1.71 (s, 3H), 1.55-1.62 (m, 2H), 1.43-1.46 (m, 2H), 1.03 (s, 6H).

Step 2:

To a solution of chlorohydrin 85 (0.22 g, 0.69 mmol) in DMF (5 mL) wasadded potassium phthalimide (0.128 g, 0.69 mmol) and the mixture washeated at 60° C. for 18 h. After cooling to room temperature, themixture was partitioned between MTBE and water. The combined organicswere washed with water, 5% aqueous LiCl and brine, dried over Na₂SO₄ andconcentrated under reduced pressure. Purification by flashchromatography (0-5% then 30% EtOAc-heptane) provided epoxide 86 as acolorless oil. Yield (0.07 g, 36%): ¹H NMR (400 MHz, CDCl₃) δ 7.26-7.33(m, 3H), 7.12 (d, J=6.9 Hz, 1H), 6.69 (dd, J=16.2, 0.8 Hz, 1H), 6.34 (d,J=16.4 Hz, 1H), 3.16-3.20 (m, 1H), 2.93 (dd, J=14.4, 5.6 Hz, 1H),2.80-2.84 (m, 2H), 2.59 (dd, J=4.8, 2.8 Hz, 1H), 2.05 (t, J=6.0 Hz, 2H),1.78 (s, 3H), 1.62-1.70 (m, 2H), 1.49-1.52 (m, 2H), 1.08 (s, 6H).

Step 3:

To a solution of epoxide 86 (0.07 g, 0.25 mmol) in DMF (5 mL) was addedNaN₃ (0.032 g, 0.49 mmol) and the mixture was heated at 65° C. for 15 h.Additional NaN₃ (0.032 g, 0.49 mmol) was added and the mixture washeated at 85° C. for 2 h. After cooling to room temperature, the mixturewas partitioned between EtOAc and water. The combined organics werewashed with water, 5% aqueous LiCl and brine, dried over Na₂SO₄ andconcentrated under reduced pressure to give azide 87. This material wastaken on to the next synthetic step without further purification. Yield(0.07 g, 87%).

Step 4:

To a solution of azide 87 in THF (3 mL) was added triphenylphosphine(0.059 g, 0.22 mmol) and water (1 mL). The reaction mixture was stirredat room temperature for 2 h then heated to reflux for 2 h. After coolingto room temperature, the mixture was partitioned between EtOAc andwater. The combined organics were washed with water and brine, driedover Na₂SO₄ and concentrated under reduced pressure. Purification byflash chromatography (20% EtOAc-hexanes then 2:28:70 concentratedaqueous NH₄OH:EtOH:CH₂Cl₂) gave Example 66 as a colorless oil. Yield(0.0235 g, 31% for two steps): ¹H NMR (400 MHz, DMSO-d₆) δ 7.19-7.28 (m,3H), 7.06 (d, J=7.6 Hz, 1H), 6.68 (d, J=16.4 Hz, 1H), 6.29 (d, J=16.4Hz, 1H), 4.75 (br s, 1H), 4.48 (br s, 2H), 3.62-3.70 (m, 1H), 2.58-2.71(m, 4H), 2.00 (t, J=6.0 Hz, 2H), 1.71 (s, 3H), 1.57-1.60 (m, 2H),1.43-1.46 (m, 2H), 1.03 (s, 6H).

Example 67 Preparation of(E)-2-fluoro-3-(3-((E/Z)-2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)prop-2-en-1-amine

(E)-2-Fluoro-3-(34(E/Z)-2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)prop-2-en-1-aminewas prepared according to Scheme 30.

Step 1:

A solution of ethyl 2-(diethoxyphosphoryl)-2-fluoroacetate (0.5675 g,2.34 mmol) in THF (5 mL) was added to a mixture of NaH (0.0652 g, 2.72mmol) in THF (3 mL). The reaction mixture was stirred at roomtemperature for 10 min then cooled to −78° C. A solution of aldehyde 69(0.3373 g, 1.33 mmol) in THF (7 mL) was added and the mixture wasstirred for 1 h, 45 min. The mixture was allowed to warm to roomtemperature then heated at 60° C. overnight. A solution of ethyl2-(diethoxyphosphoryl)-2-fluoroacetate (0.5 mL, 2.47 mmol) and NaH(0.0425 g, 1.77 mmol) in THF (5 mL) was added to the reaction mixture;the temperature was then increased to 70° C. and stirred overnight.After cooling to room temperature, the mixture was concentrated underreduced pressure and partitioned between hexanes and water. The combinedorganics were washed with brine, dried over MgSO₄ and concentrated underreduced pressure to give ester 88 as an oil. This material was used inthe next synthetic step without purification. ¹H NMR (400 MHz, CDCl₃) δ7.46 (s, 1H), 7.30-7.38 (m, 3H), 6.92 (d, J=22.0 Hz, 1H), 6.69 (dd,J=16.4, 0.8 Hz, 1H), 6.33 (d, J=16.4 Hz, 1H), 4.25 (q, J=7.2 Hz, 2H),2.05 (t, J=6.4 Hz, 2H), 1.76-1.77 (m, 3H), 1.62-1.68 (m, 2H), 1.49-1.52(m, 2H), 1.23 (t, J=7.2 Hz, 3H), 1.07 (s, 6H).

Step 2:

To an ice-cold solution of ester 88 (˜1.33 mmol) in diethyl ether (20mL) was added a solution of diisobutyl aluminum hydride (DIBAL-H, 4 mLof a 1.0 M solution in THF, 4.0 mmol) The reaction was stirred at 0° C.overnight. After warming to room temperature, the mixture waspartitioned between EtOAc and water. The combined organics were washedwith saturated aqueous NH₄Cl and brine, dried over Na₂SO₄ andconcentrated under reduced pressure. Purification by flashchromatography (10 to 50% EtOAc-hexanes gradient) gave alcohol 89 as anoil. Yield (0.1069 g, 27%): ¹H NMR (400 MHz, CDCl₃) δ 7.26-7.35 (m, 3H),7.10 (d, J=6.8 Hz, 1H), 6.70 (dd, J=16.4, 1.2 Hz, 1H), 6.41 (d, J=20.0Hz, 1H), 6.33 (d, J=16.4 Hz, 1H), 4.39 (dd, J=21.6, 6.0 Hz, 2H), 2.05(t, J=6.0 Hz, 2H), 1.89 (t, J=6.4 Hz, 1H), 1.76 (s, 3H), 1.62-1.68 (m,2H), 1.48-1.51 (m, 2H), 1.07 (s, 6H).

Step 3:

To a solution of alcohol 89 (0.0969 g, 0.323 mmol), triphenylphosphine(0.1266 g, 0.48 mmol) and phthalimide (0.0580 g, 0.39 mmol) in THF (5mL) was added a solution of diethyl azodicarboxylate (0.0880 g, 0.51mmol) in THF 2 mL). The reaction was stirred at room temperature for 25min then concentrated under reduced pressure. Purification by flashchromatography (6 to 50% EtOAc-hexanes gradient) gave phthalimide 90 asan oil. Yield (0.0847 g, 61%): ¹H NMR (400 MHz, CDCl₃) δ 7.81-7.85 (m,2H), 7.70-7.73 (m, 2H), 7.51 (s, 1H), 7.26-7.31 (m, 3H), 6.77 (dd,J=16.4, 0.8 Hz, 1H), 6.45 (d, J=22.0 Hz, 1H), 6.36 (d, J=16.4 Hz, 1H),4.68 (d, J=15.6 Hz, 2H), 2.05 (t, J=6.4 Hz, 2H), 1.78 (s, 3H), 1.61-1.68(m, 2H), 1.49-1.52 (m, 2H), 1.09 (s, 6H).

Step 4:

Phthalimide 90 was deprotected according to the method used in Example32 except that the reaction was conducted at room temperature overnight.Example 67 was isolated as an oil. Yield (0.0544 g, 93%): ¹H NMR (400MHz, CDCl₃) δ 7.29-7.33 (m, 2H), 7.20 (s, 1H), 7.04 (dt, J=6.4, 1.6 Hz,1H), 6.68 (dd, J=16.4, 0.8 Hz, 1H), 6.32 (d, J=16.4 Hz, 1H), 6.25 (d,J=22.0 Hz, 1H), 3.60 (d, J=21.6 Hz, 2H), 2.04 (t, J=6.4 Hz, 2H), 1.76(s, 3H), 1.61-1.67 (m, 2H), 1.42-1.51 (m, 2H), 1.35 (br s, 2H), 1.09 (s,6H).

Example 68 Preparation of(E)-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)prop-2-yn-1-amine

(E)-3-(3-(2-(2,6,6-Trimethylcyclohex-1-enyl)vinyl)phenyl)prop-2-yn-1-aminewas prepared according to Scheme 31.

Step 1:

A solution of 2-(prop-2-ynyl)isoindoline-1,3-dione (1.85 g, 10.0 mmol)and 3-bromobenzaldehyde (1.16 mL, 10.0 mmol) in triethylamine (25 mL)and THF (25 mL) were degassed by bubbling with argon. PdCl₂(PPh₃)₂(0.350 g, 0.05 mmol), and CuI (0.095 g, 0.5 mmol) were added and themixture was degassed again then heated at 70° C. overnight. Aftercooling to room temperature, the mixture was diluted with EtOAc and thesolids were removed by filtration. The filtrate was concentrated underreduced pressure and the residue was purified by flash chromatography (7to 60% EtOAc-hexanes gradient) to give a light orange solid. Thismaterial was triturated with hexanes and the solids were removed byfiltration. The filtrate was concentrated under reduced pressure toafford alkyne 91 as a light-yellow solid. (Yield 0.533 g, 18%): ¹H NMR(400 MHz, DMSO-d₆) δ 9.98 (s, 1H), 7.88-7.96 (m, 6H), 7.76 (dt, J=7.6,1.2 Hz, 1H), 7.60 (t, J=7.6 Hz, 1H), 4.68 (s, 2H).

Step 2:

Alkyne 91 was coupled with phosphonium salt 24 according to the methodused in Example 44 except that the aldehyde was added neat and thereaction was stirred at room temperature for 30 min. The reactionmixture was partitioned between EtOAc and water and the combinedorganics were washed with water, dried over Na₂SO₄ and concentratedunder reduced pressure. Purification by flash chromatography (7 to 60%EtOAc-hexanes gradient) gave alkene 92 as a light yellow foam. Yield(0.342 g, 83%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.84-7.92 (m, 4H), 7.53 (s,1H), 7.51 (d, J=7.6 Hz, 1H), 7.23-7.31 (m, 2H), 6.77 (d, J=16.4 Hz, 1H),6.33 (d, J=16.8 Hz, 1H), 4.62 (s, 2H), 1.99 (t, J=5.6 Hz, 2H), 1.68 (s,3H), 1.52-1.59 (m, 2H), 1.41-1.44 (m, 2H), 1.00 (s, 6H).

Step 3:

Alkene 92 was deprotected according to the method used in Example 32except that the reaction was heated at reflux for 3.5 h. After coolingto room temperature, the solids were removed from the mixture byfiltration. The filtrate was concentrated under reduced pressure. Theresidue was suspended in hexanes and sonicated then solids were removedby filtration through Celite. The filtrate was concentrated underreduced pressure and the sonication/filtration procedure was repeated.Example 68 was isolated as an oil. Yield (0.196 g, 83%): ¹H NMR (400MHz, DMSO-d₆) δ 7.43-7.47 (m, 2H), 7.29 (t, J=7.6 Hz, 1H), 7.20-7.23 (m,1H), 6.74 (dd, J=16.4, 0.8 Hz, 1H), 6.32 (d, J=16.4 Hz, 1H), 3.48 (s,2H), 2.00 (t, J=6.4 Hz, 1H), 1.79 (br s, 2H), 1.70 (s, 3H), 1.54-1.62(m, 2H), 1.43-1.46 (m, 2H), 1.03 (s, 6H).

Example 69 Preparation of (E)-3-(3-(non-4-en-5-yl)phenyl)propan-1-amine

(E)-3-(3-(Non-4-en-5-yl)phenyl)propan-1-amine was prepared according toScheme 32.

Step 1:

To a solution of 3-(3-bromophenyl)propionic acid (93) (5.0 g, 21.8 mmol)and N-hydroxysuccinimide (2.51 gr., 21.8 mmole) in CH₂Cl₂ (100 ml) wasadded dicyclohexylcarbodiimide (4.50 g, 21.8 mmol) and the mixture wasstirred at room temperature for 45 min. The precipitate was removed byfiltration and the filtrate was cooled in an ice bath. Ammonia gas wasbubbled into the solution for 2 min then allowed to warm to roomtemperature overnight. The reaction mixture was concentrated underreduced pressure and the residue was dissolved in EtOAc. This solutionwas washed with saturated aqueous NaHCO₃ and brine, dried over Na₂SO₄and concentrated under reduced pressure to an oil which was trituratedwith hexanes. The product was collected by filtration to give3-(3-bromophenyl)propionamide (94) as a white solid. This material wastaken on to the next synthetic step without further purification. Yield(4.62 g, 93%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.40 (s, 1H), 7.35 (dt,J=6.4, 2.4 Hz, 1H), 7.26 (br s, 1H), 7.18-7.24 (m, 2H), 6.75 (br s, 1H),2.78 (t, J=7.6 Hz, 2H), 2.34 (t, J=7.6 Hz, 2H).

Step 2:

To a solution of 3-(3-bromophenyl)propionamide (94) (4.5 g, 19.7 mmol)in THF (50 ml) under argon was added BH₃-THF complex (39.4 mL of a 1.0 Msolution in THF, 39.4 mmol) and the mixture was stirred at roomtemperature for 18 h. The reaction was quenched with the cautiousaddition of 6 M HCl to pH 1. After stirring 4 h the solution wasadjusted to pH>10 with the addition of 50% aqueous NaOH. This aqueoussolution was extracted with EtOAc. The combined organics were washedwith brine, dried over Na₂SO₄ and concentrated under reduced pressure togive 3-(3-bromophenyl)propan-1-amine (95) as an oil. This material wasused in the next synthetic step without purification. ¹H NMR (400 MHz,DMSO-d₆) δ 7.40 (s, 1H), 7.35 (dt, J=6.4, 2.4 Hz, 1H), 7.26 (br s, 1H),7.18-7.24 (m, 2H), 6.75 (br s, 1H), 2.78 (t, J=7.6 Hz, 2H), 2.34 (t,J=7.6 Hz, 2H).

Step 3:

To a solution of amine 95 (˜19.7 mmol) in THF (50 mL) was addeddi-tert-butyl dicarbonate (4.74 g, 21.7 mmol) and the mixture wasstirred at room temperature for 3 h. The mixture was then concentratedunder reduced pressure and purified by flash chromatography (30% diethylether-hexanes) to give aryl bromide 96 as an oil. (Yield 3.62 g, 58% fortwo steps): ¹H NMR (400 MHz, DMSO-d₆) δ 7.33 (s, 1H), 7.26-7.34 (m, 1H),7.08-7.20 (m, 2H), 4.55 (br s, 1H), 3.15 (q, J=6 Hz, 2H), 2.61 (t, J=8.0Hz, 2H), 1.79 (quint, J=7.6 Hz, 2H), 1.44 (s, 9H).

Step 4:

To a −78° C. solution of aryl bromide 96 (0.650 g, 2.07 mmol, crude) inanhydrous THF (20 mL) was added MeLi (1.36 mL of a 1.6 M solution indiethyl ether, 2.17 mmol) and the mixture was stirred for 10 mintert-Butyl lithium (2.5 mL of a 1.7 M solution in pentane, 4.24 mmol)was added and the reaction mixture was stirred at −78° C. for 45 minfollowed by the addition of 5-nonanone (0.324 g, 2.28 mmol) Afterallowing the mixture to warm to room temperature, the reaction wasquenched with the addition of saturated aqueous NH₄Cl (15 mL) and the pHwas adjusted to 5 with 1M HCl. The mixture was extracted with EtOAc andthe combined organics were dried over Na₂SO₄ and concentrated underreduced pressure to give alcohol 97 as an oil. Yield (0.090 g, 12%): ¹HNMR (400 MHz, DMSO-d₆) δ 7.14-7.19 (m, 3H), 6.97 (d, J=8.0 Hz, 1H), 6.87(t, J=4.0 Hz, 1H), 4.48 (s, 1H), 2.92 (q, J=8.0 Hz, 2H), 2.53 (t, J=8.0Hz, 2H), 1.59-1.74 (m, 6H), 1.37 (s, 9H), 1.15-1.23 (m, 6H), 0.84-0.91(m, 2H), 0.77 (t, J=8.0 Hz. 6H).

Step 5:

A solution of alcohol 97 (0.081 g, 0.215 mmol) in HCl (2 mL of a 4.2 Msolution in EtOAc, 8.4 mmol) was stirred at room temperature overnightthen concentrated under reduced pressure. Example 69 hydrochloride wasobtained as an oil. (Yield 0.066 g, quant.): ¹H NMR (400 MHz, DMSO-d₆) δ7.94 (br s, 3H), 7.11-7.37 (m, 3H), 7.07 (d, J=8.0 Hz, 1H), 5.63 (t,J=8.0 Hz, 1H), 2.77-2.80 (m, 2H), 2.64 (t, J=8.0 Hz, 2H), 2.47 (t, J=8.0Hz, 2H), 2.15 (q, J=8.0 Hz, 2H), 1.81-1.91 (m, 2H), 1.44 (q, J=8.0 Hz,2H), 1.17-1.27 (m, 4H), 0.93 (t, J=8.0 Hz, 3H), 0.83 (t, J=8.0 Hz, 3H).

Example 70 Preparation of (E)-2-(3-(3-aminopropyl)styryl)phenol

(E)-2-(3-(3-Aminopropyl)styryl)phenol was prepared according to Scheme15 with modifications.

Step 1:

2-Hydroxybenzyltriphenylphosphonium bromide was coupled with phthalimide29 according to the method used in Example 45 except that the reactionwas stirred at room temperature overnight. The reaction mixture was thenconcentrated under reduced pressure and partitioned between EtOAc andwater. The combined organics were washed with saturated aqueous NH₄Cland water, dried over Na₂SO₄ and concentrated under reduced pressure.Purification by flash chromatography (5 to 60% EtOAc-hexanes gradient)gave (E)-2-(3-(3-(2-hydroxystyryl)phenyl)propyl)isoindoline-1,3-dione asa light yellow foam. Yield (0.165 g, 43%): ¹H NMR (400 MHz, DMSO-d₆) δ9.69 (s, 1H), 7.78-7.84 (m, 4H), 7.53 (dd, J=8.0, 1.6 Hz, 1H), 7.28-7.38(m, 3H), 7.22 (t, J=8.4 Hz, 1H), 7.05-7.14 (m, 3H), 6.76-6.86 (m, 2H),3.61 (t, J=6.8 Hz, 2H), 2.63 (t, J=7.2 Hz, 2H), 1.89-1.97 (m, 2H).

Step 2:

(E)-2-(3-(3-(2-Hydroxystyryl)phenyl)propyl)isoindoline-1,3-dione wasdeprotected with hydrazine according to the method used in Example 68.After stirring overnight, the reaction mixture was concentrated underreduced pressure and the residue was suspended in 20% EtOAc-hexanes andsonicated. Solids were removed by filtration and the filtrate wasconcentrated under reduced pressure. Purification by flashchromatography (15% 7 M NH₃ in MeOH-EtOAc) gave Example 70 as a yellowfoam. Yield (0.736 g, 69%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.54 (dd, J=8.0,1.6 Hz, ¹H), 7.31-7.39 (m, 3H), 7.24 (t, J=8.4 Hz, 1H), 7.15 (d, J=16.4Hz, 1H), 7.05-7.09 (m, 2H), 6.85 (dd, J=8.0, 1.6 Hz, 1H), 6.79 (ddd,J=7.6, 0.8 Hz, 1H), 2.60 (t, J=6.8 Hz, 2H), 2.54 (t, J=7.2 Hz, 2H),1.60-1.67 (m, 2H).

Example 71 Preparation of(E)-3-(5-(2,6-dichlorostyryl)-2-methoxyphenyl)propan-1-amine

(E)-3-(5-(2,6-Dichlorostyryl)-2-methoxyphenyl)propan-1-amine wasprepared according to Scheme 33.

Step 1:

Aryl aldehyde 98 was protected according the method used in Example 32except that the reaction mixture was stirred at room temperature for 1.5h. Dimethyl acetal 99 was isolated as an oil. Yield (6.89 g, quant.): ¹HNMR (400 MHz, CDCl₃) δ 7.64 (d, J=2.0 Hz, 1H), 7.34 (dd, J=8.4, 2.0 Hz,1H), 6.88 (d, J=8.4 Hz, 1H), 5.33 (s, 1H), 3.90 (s, 3H), 3.30 (s, 6H).

Step 2:

Dimethyl acetal 99 was coupled with N-allyl-2,2,2-trifluoroacetamideaccording to the method used in Example 45 except that the reaction washeated at 70° C. overnight. After cooling to room temperature, themixture was concentrated under reduced pressure. The residue wassuspended in EtOAc and the solids were removed by filtration. Thefiltrate was concentrated under reduced pressure and purified by flashchromatography (10 to 50% EtOAc-hexanes gradient) to give alkene 100 asa yellow oil which solidified upon standing under vacuum. (Yield (1.82g, 56%): ¹H NMR (400 MHz, CDCl₃) δ 7.48 (d, J=2.0 Hz, 1H), 7.32 (dd,J=8.4, 2.0 Hz, 1H), 6.91 (d, J=16.0 Hz, 1H), 6.87 (d, J=8.8 Hz, 1H),6.40 (br s, 1H), 6.25 (dt, J=16.0, 6.8 Hz, 1H), 5.34 (s, 1H), 4.14 (t,J=6.8 Hz, 2H), 3.86 (s, 3H), 3.31 (s, 6H).

Step 3:

To a degassed solution of alkene 100 (0.8469 g, 2.54 mmol) in EtOH(abs., 15 mL) was added 10% Pd/C (0.0419 g). The mixture was placedunder a H₂ atmosphere and stirred at room temperature for 3.5 h. Solidswere removed by filtration and the filtrate was concentrated underreduced pressure to give crudeN-(3-(5-(dimethoxymethyl)-2-methoxyphenyl)propyl)-2,2,2-trifluoroacetamide.This material was used in the next synthetic step without purification.

Step 4:

N-(3-(5-(dimethoxymethyl)-2-methoxyphenyl)propyl)-2,2,2-trifluoroacetamidewas deprotected according to the method used in Example 32 except thatthe reaction was stirred for 1.5 h. Trifluoroacetamide 101 was isolatedas white crystals. This material was used in the next synthetic stepwithout purification. Yield (0.7118 g, 97% for two steps): ¹H NMR (400MHz, DMSO-d₆) δ 9.83 (s, 1H), 9.42 (br s, 1H), 7.78 (dd, J=8.4, 2.0 Hz,1H), 7.69 (d, J=8.4 Hz, 1H), 7.15 (d, J=8.4 Hz, 1H), 3.87 (s, 3H), 3.18(q, J=6.8 Hz, 2H), 2.60 (t, J=7.6 Hz, 2H), 1.71-1.79 (m, 2H).

Step 5:

Trifluoroacetamide 101 was coupled to phosphonium salt 55 according tothe method used in Example 46 except that it was stirred at −78° C.overnight. After warming to room temperature, the reaction was heated toreflux for 2 h. The reaction mixture was cooled to room temperature thenconcentrated under reduced pressure. The residue was suspended in 5%EtOAc-heptane and sonicated. The solids were removed by filtration andthe filtrate was concentrated under reduced pressure. Purification byflash chromatography (10 to 40% EtOAc-hexanes gradient) afforded alkene102 as an oil. Yield (0.4765 g, 75%): ¹H NMR (400 MHz, CDCl₃) δ 7.39(dd, J=8.4, 2.0 Hz, 1H), 7.33-7.35 (m, 3H), 7.07-7.11 (m, 2H), 6.98 (d,J=16.4 Hz, 1H), 6.89 (d, J=8.8 Hz, 1H), 6.73 (br s, 1H), 3.88 (s, 3H),3.33 (q, J=6.8 Hz, 2H), 2.74 (t, J=7.6 Hz, 2H), 1.87-1.94 (m, 2H).

Step 6:

To a solution of alkene 102 (0.0488 g, 0.113 mmol) in MeOH—H₂O (5:1, 2mL) was added K₂CO₃ (0.0352, 0.26 mmol). The mixture was stirred at roomtemperature for 20 min then EtOH (2 mL) was added and the mixture wasstirred overnight. The reaction mixture was concentrated under reducedpressure and purified by flash chromatography (1:5:5 7 M NH₃ in MeOH:EtOAc: hexanes) to give Example 71 as an oil. Yield (0.0291 g, 77%): ¹HNMR (400 MHz, CDCl₃) δ 7.32-7.37 (m, 4H), 7.05-7.12 (m, 2H), 6.98 (d,J=16.4 Hz, 1H), 6.85 (d, J=8.8 Hz, 1H), 3.85 (s, 3H), 2.74 (t, J=7.6 Hz,2H), 2.68 (t, J=7.6 Hz, 2H), 1.73-1.80 (m, 2H), 1.27 (br s, 2H).

Example 72 Preparation of(E)-3-amino-2-methyl-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-one

(E)-3-Amino-2-methyl-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-onewas prepared according to Scheme 22 with modifications.

Step 1:

(E)-3-Amino-2-methyl-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-ol(Example 59) was reacted with di-tert-butyl dicarbonate according themethod used in Example 55 to give (E)-tert-butyl3-hydroxy-2-methyl-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propylcarbamateas an oil. Yield (0.2728 g, 39%).

Step 2:

To a solution of (E)-tert-butyl3-hydroxy-2-methyl-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propylcarbamate(0.2728 g, 0.64 mmol) in CH₂Cl₂ (10 mL) under argon was added MnO₂(2.4751 g, 28.5 mmol) The mixture was stirred at room temperatureovernight. The solids were removed by filtration and the filtrate wasconcentrated under reduced pressure. Purification by flashchromatography (10 to 50% EtOAc-hexanes gradient) gave (E)-tert-butyl2-methyl-3-oxo-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propylcarbamateas an oil. (Yield 0.1333 g, 49%): ¹H NMR (400 MHz, CDCl₃) δ 7.96 (s,1H), 7.79 (d, J=7.6 Hz, 1H), 7.61 (d, J=7.6 Hz, 1H), 7.41 (t, J=8.0 Hz,1H), 6.75 (dd, J=16.4, 0.8 Hz, 1H), 6.38 (d, J=16.4 Hz, 1H), 4.96 (br s,1H), 3.76-3.81 (m, 1H), 3.40 (t, J=6.0 Hz, 2H), 2.04 (t, J=6.0 Hz, 2H),1.76 (s, 3H), 1.61-1.72 (m, 2H), 1.46-1.51 (m, 2H), 1.40 (s, 9H), 1.19(d, J=7.2 Hz, 3H), 1.06 (s, 6H).

Step 3:

(E)-tert-Butyl2-methyl-3-oxo-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propylcarbamatewas deprotected according to the method used in Example 61 except thatthe reaction was stirred for 1 h. After concentration, the residue wasdissolved in EtOAc and concentrated under reduced pressure. Thisprocedure was repeated with MeOH to give Example 72 hydrochloride as anoil. Yield (0.0533 g, quant.): ¹H NMR (400 MHz, DMSO-d₆) δ 7.99 (s, 1H),7.95 (br s, 3H), 7.82 (dd, J=8.0, 1.2 Hz, 2H), 7.51 (t, J=8.0 Hz, 1H),6.83 (d, J=16.4, 1H), 6.46 (d, J=16.4 Hz, 1H), 3.92-3.98 (m, 1H), 3.18(dd, J=12.8, 7.6 Hz, 1H), 2.89 (dd, J=12.8, 7.6 Hz, 1H), 2.02 (t, J=6.0Hz, 2H), 1.73 (s, 3H), 1.56-1.62 (m, 2H), 1.44-1.47 (m, 2H), 1.16 (d,J=7.2 Hz, 3H), 1.04 (s, 6H).

Example 73 Preparation of(E)-3-amino-2-fluoro-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-one

(E)-3-Amino-2-fluoro-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-onewas prepared according to Scheme 34.

Step 1:

To a −78° C. solution of lithium diisopropylamide (1 mL of a 2 Msolution in THF:heptane:ethyl benzene, 2.0 mmol) in THF (1 mL) was addeda solution of ketone 65 (0.2855 g, 0.72 mmol) in THF (6 mL) slowly. Thereaction mixture was stirred at −78° C. for 7 min, removed from thecooling bath for 6 min then cooled to −78° C. again. A solution ofN-fluorobenzenesulfonimide (0.2750 g, 0.87 mmol) in THF (5 mL) was addeddropwise slowly and the mixture was stirred at −78° C. for 25 min. Thecooling bath was removed and the reaction was quenched with the additionof saturated aqueous NH₄Cl. The mixture was extracted with EtOAc and thecombined organics were washed with brine, dried over MgSO₄ andconcentrated under reduced pressure. Purification by flashchromatography (10 to 40% EtOAc-hexanes gradient) gave fluoride 103 as agummy yellow solid. Yield (0.1634 g, 55%): ¹H NMR (400 MHz, CDCl₃) δ8.05 (s, 1H), 7.85 (d, J=7.6 Hz, 1H), 7.63 (d, J=7.6 Hz, 1H), 7.44 (t,J=8.0 Hz, 1H), 6.79 (d, J=16.4, 1H), 6.38 (d, J=16.4 Hz, 1H), 5.73-5.86(m, 1H), 5.04 (br s, 1H), 3.80-3.95 (m, 1H), 3.40-3.52 (m, 1H), 2.04 (t,J=6.0 Hz, 2H), 1.75 (s, 3H), 1.55-1.67 (m, 2H), 1.47-1.52 (m, 2H), 1.43(s, 9H), 1.06 (s, 6H).

Step 2:

Fluoride 103 was deprotected according to the method used in Example 72to give Example 73 hydrochloride as an oil. Yield (0.0481 g, quant.): ¹HNMR (400 MHz, DMSO-d₆) δ 8.32 (br s, 3H), 8.00 (s, 1H), 7.87 (d, J=8.0Hz, 1H), 7.81 (d, J=8.0 Hz, 1H), 7.54 (t, J=8.0 Hz, 1H), 6.85 (d,J=16.4, 1H), 6.46 (d, J=16.4 Hz, 1H), 6.32 (ddd, J=47.6, 8.4, 2.8 Hz,1H), 3.24-3.50 (m, 2H), 2.02 (t, J=6.0 Hz, 2H), 1.73 (s, 3H), 1.56-1.62(m, 2H), 1.45-1.47 (m, 2H), 1.05 (s, 6H).

Example 74 Preparation of(R,E)-1-amino-3-(3-(2,6-dichlorostyryl)phenyl)propan-2-ol

(R,E)-1-Amino-3-(3-(2,6-dichlorostyryl)phenyl)propan-2-ol was preparedfrom aryl bromide 56 according to Scheme 29 with modifications.

Step 1:

To a −78° C. solution of aryl bromide 56 (2.01 g, 6.13 mmol) in THF (16mL) was added n-butyl lithium (3.8 mL of a 1.6 M solution in hexanes,6.1 mmol) dropwise over 11 min. The mixture was stirred for 7 min thenBF₃-diethyl etherate (0.7 mL, 5.5 mmol) was added dropwise. Afterstirring for 3 min, a solution of (R)-epichlorohydrin (0.38 mL, 4.8mmol) in THF (4 mL) was added dropwise over 12 min. The reaction mixturewas stirred for 1 h, 15 min at −78° C. then a second aliquot ofBF₃-diethyl etherate (0.23 mL, 1.84 mmol) and (R)-epichlorohydrin (0.096mL, 1.23 mmol) were added. The mixture was stirred for 15 min at −78° C.then additional BF₃-diethyl etherate (0.23 mL, 1.84 mmol) and(R)-epichlorohydrin (0.096 mL, 1.23 mmol) were added. The mixture wasstirred for 30 min at −78° C. then quenched with water. After warming toroom temperature, the mixture was partitioned between MTBE and water.The combined organics were washed with water and brine, dried overNa₂SO₄ and concentrated under reduced pressure. Purification by flashchromatography (1:6 EtOAc: heptane) gave(R,E)-1-chloro-3-(3-(2,6-dichlorostyryl)phenyl)propan-2-ol. Yield (0.67g, 32%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.51 (d, J=8.0 Hz, 2H), 7.45 (d,J=7.6 Hz, 2H), 7.29-7.33 (m, 2H), 7.20 (d, J=7.6 Hz, 1H), 7.13 (d,J=16.6 Hz, 1H), 7.05 (d, J=16.6 Hz, 1H), 5.19 (d, J=5.6 Hz, 1H),3.85-3.95 (m, 1H), 3.57 (dd, J=11.2, 4.8 Hz, 1H), 3.49 (dd, J=11.2, 5.6Hz, 1H), 2.84 (dd, J=13.2, 4.8 Hz, 1H), 2.69 (dd, J=13.6, 8.0 Hz, 1H).

Step 2:

To a solution of(R,E)-1-chloro-3-(3-(2,6-dichlorostyryl)phenyl)propan-2-ol (0.67 g, 1.96mmol) in DMF (20 mL) was added NaI (˜25 mg, 0.16 mmol) and NaN₃ (0.64 g,9.8 mmol) and the reaction mixture was heated at 75° C. overnight. Aftercooling to room temperature, the mixture was partitioned between EtOAcand water. The combined organics were washed with water, 5% aqueous LiCland brine, filtered through Na₂SO₄ and concentrated under reducedpressure to give(R,E)-1-azido-3-(3-(2,6-dichlorostyryl)phenyl)propan-2-ol. This materialwas used in the next synthetic step without purification.

Step 3:

(R,E)-1-Azido-3-(3-(2,6-dichlorostyryl)phenyl)propan-2-ol was reducedaccording to the method used in Example 66 except that it was heated at50° C. overnight. After cooling to room temperature, the mixture waspartitioned between CH₂Cl₂ and saturated aqueous NaHCO₃. The combinedorganics were washed with brine, filtered through Na₂SO₄ andconcentrated under reduced pressure. Purification by flashchromatography (CH₂Cl₂ then 1:14:85 conc. aq. NH₄OH:EtOH:CH₂Cl₂) gaveExample 74 as a colorless oil. Yield (0.51 g, 81% for two steps): ¹H NMR(400 MHz, CDCl₃) δ 7.42 (d, J=7.6 Hz, 1H), 7.38 (s, 1H), 7.34 (d, J=8.0Hz, 2H), 7.31 (t, J=7.4 Hz, 1H), 7.17 (d, J=7.2 Hz, 1H), 7.08-7.12 (m,3H), 3.80-3.86 (m, 1H), 2.90 (dd, J=12.8, 3.6 Hz, 1H), 2.77-2.79 (m,2H), 2.64 (dd, J=12.8, 8.4 Hz, 1H), 2.28 (br s, 3H). Chiral HPLC: 99.0%major enantiomer (AUC), t_(R)=16.354 min (minor enantiomer: 1.0%,t_(R)=15.024 min).

Example 75 Preparation of(S,E)-1-amino-3-(3-(2,6-dichlorostyryl)phenyl)propan-2-ol

(S,E)-1-Amino-3-(3-(2,6-dichlorostyryl)phenyl)propan-2-ol was preparedaccording to the method used in Example 74.

Step 1:

Aryl bromide 56 was coupled with (S)-epichlorohydrin according to themethod used in Example 74 except that after the addition of a singlealiquot of (S)-epichlorohydrin the reaction was warmed to 0° C. andstirred for 12 min. After quench (at −78° C.) and extractive workup,purification by flash chromatography (1:20 EtOAc:heptane to 1:6EtOAc:heptane) gave(S,E)-1-chloro-3-(3-(2,6-dichlorostyryl)phenyl)propan-2-ol. Yield (2.04g, 22%): the ¹H NMR data was consistent with data reported above.

Step 2:

(S,E)-1-Chloro-3-(3-(2,6-dichlorostyryl)phenyl)propan-2-ol was reactedwith NaN₃ following the method used in Example 74 to give(S,E)-1-azido-3-(3-(2,6-dichlorostyryl)phenyl)propan-2-ol. This materialwas used in the next synthetic step without purification.

Step 3:

(S,E)-1-Azido-3-(3-(2,6-dichlorostyryl)phenyl)propan-2-ol was reducedand purified according to the method used in Example 74 to give Example75 as a colorless oil. Yield (1.25 g, 67% for two steps): ¹H NMR (400MHz, DMSO-d₆) δ 7.52 (d, J=8.4 Hz, 2H), 7.42-7.44 (m, 2H), 7.27-7.32 (m,2H), 7.18 (d, J=7.4 Hz, 1H), 7.11 (d, J=16.6 Hz, 1H), 7.04 (d, J=16.6Hz, 1H), 3.54-3.60 (m, 1H), 2.72 (dd, J=13.2, 4.8 Hz, 1H), 2.58 (dd,J=13.6, 8.0 Hz, 1H), 2.52 (dd, J=12.8, 4.4 Hz, 1H), 2.42 (dd, J=12.4,6.4 Hz, 1H). Chiral HPLC: 98.5% major enantiomer (AUC), t_(R)=15.024 min(minor enantiomer: 1.5%, t_(R)=16.354 min).

Example 76 Preparation of(E/Z)-(3-(3-(2,6-diethoxystyryl)phenyl)propan-1-amine

(E/Z)-(3-(3-(2,6-Diethoxystyryl)phenyl)propan-1-amine (isomer ratio69:31 trans: cis) was prepared according to the method used in Example32.

Step 1:

To an ice cold solution of 2,6-diethoxybenzyl alcohol (1.0 g, 5.2 mmol)in THF (10 mL) was added phosphorous tribromide (0.48 mL, 5.1 mmol)dropwise. The reaction was allowed to warm to room temperature andstirred for 1 h. After quenching the reaction with water, the mixturewas extracted with EtOAc. The combined organics were washed with brineand dried over Na₂SO₄. The solution was concentrated under reducedpressure to give 2,6-diethoxybenzyl bromide as a brown oil. Yield (1.3g, 98%).

Step 2:

To a solution of triphenylphosphine (1.31 g, 5.01 mmol) in toluene (6.5mL) was added 2,6-diethoxybenzyl bromide (1.3 g, 5.01 mmol) and themixture stirred at room temperature overnight. Diethyl ether was addedand the solid was collected by filtration to give(2,6-diethoxybenzyl)triphenylphosphonium bromide as an off-white solid.Yield (1.58 g, 60%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.83-7.87 (m, 3H), 7.68(ddd, J=7.6, 7.6, 3.2 Hz, 6H), 7.55 (d, J=7.6 Hz, 3H), 7.52 (d, J=7.2Hz, 3H), 7.18 (ddd, J=8.4, 8.4, 2.4 Hz, 1H), 6.45 (d, J=8.4 Hz, 2H),4.66 (d, J=14.0 Hz, 2H), 3.65 (q, J=6.8 Hz, 4H), 1.01 (t, J=6.8 Hz, 6H).

Step 2:

To an ice-cold mixture of phthalimide 29 (0.440 g, 1.5 mmol) in THF (25mL) was added (2,6-diethoxybenzyl)triphenylphosphonium bromide (0.860 g,1.65 mmol) and potassium tert-butoxide (0.336 g, 3.0 mmol) portionwise.The reaction was allowed to warm to room temperature and stirred for 2h. The mixture was then quenched with water and extracted with EtOAc.The combined organics were washed with brine, dried over Na₂SO₄ andconcentrated under reduced pressure. Purification by flashchromatography (7% EtOAc-hexanes) gave(E)-2-(3-(3-(2,6-diethoxystyryl)phenyl)propyl)isoindoline-1,3-dione as abrown oil. Yield (0.055 g, 8%): ¹H NMR (400 MHz, CDCl₃) δ 7.80-7.83 (m,2H), 7.67-7.70 (m, 2H), 7.62 (d, J=16.8 Hz, 1H), 7.45 (d, J=16.8 Hz,1H), 7.29-7.31 (m, 1H), 7.22 (t, J=7.6 Hz, 1H), 7.17 (d, J=6.4 Hz, 1H),7.11 (t, J=8.4 Hz, 1H), 7.06 (d, J=7.2 Hz, 1H), 6.60 (d, J=8.4 Hz, 2H),4.11 (dt, J=7.2, 6.8 Hz, 4H), 3.78 (t, J=7.2 Hz, 2H), 2.71 (t, J=7.6 Hz,2H), 2.03-2.11 (m, 2H), 1.50 (t, J=7.2 Hz, 6H).

Step 3:

To a solution of(E)-2-(3-(3-(2,6-diethoxystyryl)phenyl)propyl)isoindoline-1,3-dione(0.375 g, 0.824 mmol) in EtOH (5 mL) was added hydrazine hydrate (0.15mL, 2.5 mmol) The mixture was heated to reflux for 1 h. After cooling toroom temperature, diethyl ether was added and the white precipitate wasremoved by filtration. The filtrate was concentrated under reducedpressure to give Example 76 as a yellow oil. Yield (0.220 g, 82%):Trans-/cis-isomer ratio 2.2:1. Trans-isomer: ¹H NMR (400 MHz, CDCl₃) δ7.66 (d, J=16.8 Hz, 1H), 7.47 (d, J=16.8 Hz, 1H), 7.04-7.37 (m, 5H),6.56 (d, J=8.4 Hz, 2H), 4.10 (q, J=7.2 Hz, 4H), 2.65-2.78 (m, 4H),1.75-1.84 (m, 2H), 1.49 (t, J=6.8 Hz, 6H).

Example 77 Preparation of (E)-3-(3-(2-ethoxystyryl)phenyl)propan-1-amine

(E)-3-(3-(2-Ethoxystyryl)phenyl)propan-1-amine was prepared according tothe method used in Example 32.

Step 1:

Phthalimide 29 was coupled with 2-ethoxybenzyltriphenylphosphoniumbromide according to the method used in Example 76 except that thereaction was stirred at room temperature for 1 h. Purification by flashchromatography (10% EtOAc-hexanes) gave(E/Z)-2-(3-(3-(2-ethoxystyryl)phenyl)propyl)isoindoline-1,3-dione ayellow solid. Yield (0.258 g, 61%).

Step 2:

(E/Z)-2-(3-(3-(2-ethoxystyryl)phenyl)propyl)isoindoline-1,3-dione wasdeprotected following the method used in Example 76. Purification byPreparative HPLC (Method 2) gave Example 77 trifluoroacetate. Yield(0.0.035 g, 20%): 93% trans-isomer. Trans-isomer: ¹H NMR (400 MHz,CDCl₃) δ 7.91 (br s, 3H), 7.55 (dd, J=7.6, 1.2 Hz, 1H), 7.45 (d, J=16.4Hz, 1H), 7.38 (d, J=8.0 Hz, 1H), 7.13-7.22 (3H), 7.09 (d, J=16.4 Hz,1H), 7.00 (d, J=7.6 Hz, 1H), 6.92-6.95 (m, 1H), 6.87 (d, J=8.4 Hz, 1H),4.08 (dt, J=7.2, 6.8 Hz, 2H), 2.82-2.87 (m, 2H), 2.65 (t, J=7.6 Hz, 2H),1.93-2.02 (m, 2H), 1.45 (t, J=7.2 Hz, 3H).

Example 78 Preparation of(E/Z)-3-(3-(2-isopropoxystyryl)phenyl)propan-1-amine

(E/Z)-3-(3-(2-Isopropoxystyryl)phenyl)propan-1-amine was preparedaccording to the method used in Example 76.

Step 1:

2-Isopropoxybenzyltriphenylphosphonium bromide was prepared from2-isopropoxylbenzyl bromide according to the method used in Example 76,except that 1.3 equivalents of triphenylphosphine was used.2-Isopropoxybenzyltriphenylphosphonium bromide was isolated as a whitesolid. Yield (11.2 g, 99%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.86-7.89 (m,3H), 7.70 (dt, J=8.0, 3.2 Hz, 6H), 7.54-7.59 (m, 6H), 7.23-7.28 (m, 1H),6.91 (dt, J=7.6, 2.0 Hz, 1H), 6.88 (d, J=8.4 Hz, 1H), 6.74 (t, J=7.6 Hz,1H), 4.87 (d, J=14.4 Hz, 2H), 4.29 (quint, J=6.0 Hz, 1H), 0.91 (d, J=6.0Hz, 6H).

Step 2:

Phthalimide 29 was coupled with 2-isopropoxybenzyltriphenylphosphoniumbromide according to the method used in Example 76. Purification byflash chromatography (7% EtOAc-hexanes) gave(E/Z)-2-(3-(3-(2-isopropoxystyryl)phenyl)propyl)isoindoline-1,3-dione ayellow oil. Yield (0.375 g, 51%). Trans-isomer: ¹H NMR (400 MHz, CDCl₃)δ 780-7.85 (m, 2H), 7.67-7.72 (m, 2H), 7.58 (dd, J=7.6, 1.2 Hz, 1H),7.43 (d, J=16.4 Hz, 1H), 7.31 (s, 1H), 7.03-7.24 (m, 5H), 6.97-6.99 (m,1H), 6.93 (t, J=8.0 Hz, 1H), 6.87 (d, J=8.4 Hz, 1H), 4.55-4.61 (m, 1H),3.78 (t, J=7.2 Hz, 2H), 2.71 (t, J=7.6 Hz, 2H), 2.04-2.12 (m, 2H), 1.40(d, J=6.0 Hz, 6H).

Step 3:

(E/Z)-2-(3-(3-(2-isopropoxystyryl)phenyl)propyl)isoindoline-1,3-dionewas deprotected following the method used in Example 76 except that thereaction was heated at reflux for 2 h. Example 78 was isolated as ayellow oil. Yield (0.150 g, 31%): Cis-/trans-isomer ratio 4:1.Cis-isomer: ¹H NMR (400 MHz, CDCl₃) δ 7.82 (br s, 3H), 6.87-7.14 (m,8H), 6.70 (d, J=12.4 Hz, 1H), 6.55 (d, J=12.4 Hz, 1H), 4.49-4.57 (m,1H), 2.77-2.84 (m, 2H), 2.54 (t, J=7.6 Hz, 2H), 1.72-1.89 (m, 2H), 1.29(d, J=6.0 Hz, 6H)

Example 79 Preparation of(E)-4-(3-(3-aminopropyl)phenyl)-2-methylbut-3-en-2-ol

(E)-4-(3-(3-Aminopropyl)phenyl)-2-methylbut-3-en-2-ol was preparedaccording to Scheme 35.

Step 1:

Crude 3-(3-bromophenyl)propan-1-amine (95) (˜104.6 mmol) was stirredwith ethyl trifluoroacetate (30 ml) overnight. The mixture wasconcentrated under reduced pressure. Purification by flashchromatography (20% EtOAc-hexanes) gave trifluoroacetamide 104. Yield(21.1 g, 62%): ¹H NMR (400 MHz, DMSO-d₆) δ 9.40 (br s, 1H), 7.43 (s,1H), 7.36 (dt, J=7.2, 2.0 Hz, 1H), 7.19-7.25 (m, 2H), 3.16 (q, J=6.8 Hz,2H), 2.57 (t, J=7.6 Hz, 2H), 1.77 (quint, J=7.2 Hz, 2H).

Step 2:

Aryl bromide 104 was coupled to 2-methyl-3-buten-2-ol according to themethod used in Example 45 except that the reaction was heated at 100° C.overnight. The reaction mixture was partitioned between EtOAc andaqueous NH₄OAc. The combined extracts were washed with saturated aqueousNH₄OAc, saturated aqueous NaHCO₃, water and brine, dried over Na₂SO₄ andconcentrated under reduced pressure. Purification by flashchromatography (30 to 90% EtOAc-hexanes gradient) gave alkene 105 as alight yellow semi-solid. Yield (0.3881 g, 73%): ¹H NMR (400 MHz,DMSO-d₆) δ 9.41 (br s, 1H), 7.18-7.23 (m, 3H), 7.02-7.04 (m, 1H), 6.45(d, J=16.4 Hz, 1H), 6.36 (d, J=16.0 Hz, 1H), 3.17 (dt, J=7.0, 6.4 Hz,2H), 2.56 (t, J=7.2 Hz, 2H), 1.77 (app dt, J=7.6 Hz, 2H), 1.24 (s, 6H).

Step 3:

To a solution of alkene 105 (0.3608 g, 1.14 mmol) in MeOH—H₂O (8:1, 22.5mL) was added K₂CO₃ (0.37 g, 2.7 mmol) and the mixture was stirred atroom temperature for 23 h. The mixture was concentrated under reducedpressure then dissolved in ˜2% MeOH-EtOAc and solids were removed byfiltration. The filtrate was concentrated under reduced pressure and theresidue was purified by flash chromatography (90 to 100% EtOAc-hexanesgradient then 10% 7 M NH₃ in MeOH-EtOAc) to give Example 79 as acolorless oil. Yield (0.198 g, 80%): ¹H NMR (400 MHz, DMSO-d₆) δ7.19-7.23 (m, 3H), 7.04-7.06 (m, 1H), 6.54 (d, J=16.0 Hz, 1H), 6.35 (d,J=16.0 Hz, 1H), 4.65 (s, 1H), 2.61-2.66 (m, 4H), 1.74-1.81 (m, 2H), 1.59(br s, 1H), 1.37 (s, 6H).

Example 80 Preparation of(E)-3-amino-1-(3-(2,6-dichlorostyryl)phenyl)propan-1-one

(E)-3-Amino-1-(3-(2,6-dichlorostyryl)phenyl)propan-1-one was preparedaccording to Scheme 22 with modifications.

Step 1:

To a solution of (E)-3-amino-1-(3-(2,6-dichlorostyryl)phenyl)propan-1-ol(Example 50) (0.240 g, 0.745 mmol) in CH₂Cl₂ (10 mL) was addeddi-tert-butyl dicarbonate (0.2503 g, 1.15 mmol) and the mixture wasstirred at room temperature for 30 min. MnO₂ (2.1897 g, 25.2 mmol) wasthen added and the reaction stirred at room temperature overnight.Solids were removed by filtration through a pad of silica gel and thefiltrate was concentrated under reduced pressure. Purification by flashchromatography (10 to 50% EtOAc-hexanes gradient) afforded(E)-tert-butyl 3-(3-(2,6-dichlorostyryl)phenyl)-3-oxopropylcarbamate asan oil. Yield (0.0478 g, 31%): ¹H NMR (400 MHz, CDCl₃) δ 8.05 (s, 1H),7.88 (dt, J=8.0, 1.6 Hz, 1H), 7.75 (dt, J=8.0, 1.2 Hz, 1H), 7.49 (t,J=8.0 Hz, 1H), 7.35 (d, J=8.0 Hz, 2H), 7.19 (s, 2H), 7.13 (t, J=8.0 Hz,1H), 5.15 br s, 1H), 3.56 (q, J=6.0 Hz, 2H), 3.24 (q, J=6.0 Hz, 2H),1.43 (s, 9H).

Step 2:

(E)-tert-Butyl 3-(3-(2,6-dichlorostyryl)phenyl)-3-oxopropylcarbamate wasdeprotected according to the method used in Example 45 except that thereaction was stirred for 1.5 h. The precipitate was collected byfiltration and dried under vacuum to give Example 80 hydrochloride as awhite solid. Yield (0.0201 g, 50%): ¹H NMR (400 MHz, DMSO-d₆) δ 8.15 (s,1H), 7.96 (d, J=8.0 Hz, 1H), 7.92 (d, J=8.0 Hz, 1H), 7.77 (br s, 3H),7.60 (t, J=8.0 Hz, 1H), 7.55 (d, J=8.0 Hz, 2H), 7.34 (t, J=8.0 Hz, 1H),7.27 (d, J=16.8 Hz, 1H), 7.19 (d, J=16.8 Hz, 1H), 3.43 (t, J=6.4 Hz,2H), 3.12-3.20 (m, 2H).

Example 81 Preparation of(E)-1-amino-3-(3-(2,6-dichlorostyryl)phenyl)propan-2-one

(E)-1-Amino-3-(3-(2,6-dichlorostyryl)phenyl)propan-2-one was preparedaccording to the methods used in Examples 55 and 56 with modifications.

Step 1:

To a solution of(S,E)-1-amino-3-(3-(2,6-dichlorostyryl)phenyl)propan-2-ol (Example 74)(1.19 g, 3.69 mmol) in THF (25 mL) was added N, N-diisopropylethylamine(0.675 mL, 3.88 mmol) and di-tert-butyl dicarbonate (0.85 g, 3.9 mmol).Additional THF (5 mL) was added and the reaction was stirred at roomtemperature for 3 h. The mixture was partitioned between EtOAc and 5%aqueous NaHSO₄ and the combined organics were washed with 5% aqueousNaHSO₄, water, saturated aqueous NaHCO₃, water and brine then dried overNa₂SO₄. The solution was concentrated under reduced pressure to give(E)-tert-butyl 3-(3-(2,6-dichlorostyryl)phenyl)-2-hydroxypropylcarbamateas a colorless oil. This material was used in the next synthetic stepwithout purification. Yield (1.51 g, 97%): ¹H NMR (400 MHz, DMSO-d₆) δ7.52 (d, J=8.4 Hz, 2H), 7.42 (d, J=7.6 Hz, 2H), 7.27-7.32 (m, 2H), 7.17(d, J=7.6 Hz, 1H), 7.11 (d, J=16.6 Hz, 1H), 7.04 (d, J=16.8 Hz, 1H),6.69 (t, J=5.9 Hz, 1H), 4.72 (d, J=5.6 Hz, 1H), 3.66-3.74 (m, 1H),2.86-3.05 (m, 2H), 2.72 (dd, J=14.0, 4.8 Hz, 1H), 2.54 (dd, J=13.6, 8.0Hz, 1H), 1.36 (s, 9H).

Step 2:

To a solution of (E)-tert-butyl3-(3-(2,6-dichlorostyryl)phenyl)-2-hydroxypropylcarbamate (1.08 g, 2.56mmol) in CH₂Cl₂ (20 mL) was added Celite (0.8 g) and pyridiniumchlorochromate (0.661 g, 3.06 mmol) The reaction mixture was stirred atroom temperature for 1.5 h then additional Celite (0.70 g) andpyridinium chlorochromate (0.552 g, 2.56 mmol) were added. The mixturewas stirred for 1 h then the solids were removed by filtration throughCelite. The filtrate was concentrated under reduced pressure thenpurified by flash chromatography (10 to 70% EtOAc-hexanes gradient) togive (E)-tert-butyl3-(3-(2,6-dichlorostyryl)phenyl)-2-oxopropylcarbamate. Yield (0.49 g,45%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.52 (d, J=8.4 Hz, 2H), 7.48 (d, J=7.8Hz, 1H), 7.42 (s, 1H), 7.29-7.36 (m, 2H), 7.15 (d, J=7.4 Hz, 1H), 7.12(d, J=16.6 Hz, 1H), 7.04 (d, J=16.6 Hz, 1H), 7.05-7.06 (m, 1H), 3.88 (d,J=6.0 Hz, 2H), 3.78 (s, 2H), 1.36 (s, 9H).

Step 3:

To a solution of (E)-tert-butyl3-(3-(2,6-dichlorostyryl)phenyl)-2-oxopropylcarbamate 0.133 g, 0.32mmol) in EtOAc (1.5 mL) was added HCl-EtOAc (0.75 mL of a 4.2 Msolution, 3.2 mmol) and the mixture was stirred at room temperature for2 h. The white precipitate was collected by filtration and dried in avacuum oven at 45° C. to give Example 81 as a white solid. Yield (0.0695g, 62%): ¹H NMR (400 MHz, DMSO-d₆) δ 8.11 (br s, 3H), 7.51-7.54 (m, 3H),7.47 (s, 1H), 7.38 (t, J=7.6 Hz, 1H), 7.32 (t, J=8.0 Hz, 1H), 7.20 (d,J=7.6 Hz, 1H), 7.13 (d, J=16.6 Hz, 1H), 7.06 (d, J=16.6 Hz, 1H), 4.03(br s, 2H), 3.92 (s, 2H).

Example 82 Preparation of(E)-3-amino-1-(3-(2-chloro-6-methylstyryl)phenyl)propan-1-ol

(E)-3-Amino-1-(3-(2-chloro-6-methylstyryl)phenyl)propan-1-ol wasprepared according to the method used in Example 50.

Step 1:

2-Chloro-6-methylbenzaldehyde was coupled to3-bromobenzyltriphenylphosphonium bromide according to the method usedin Example 46 except that the addition of potassium tert-butoxide wasdone at −78° C. and the reaction mixture was briefly removed from thecooling bath. Purification by flash chromatography (0 to 50%EtOAc-hexanes gradient) gave(E/Z)-2-(3-bromostyryl)-1-chloro-3-methylbenzene as an oil. Yield (0.43g, 69%), trans-/cis-isomer ratio ˜1:1. Trans-isomer: ¹H NMR (400 MHz,CDCl₃) δ 7.67 (t, J=2.0 Hz, 1H), 7.40-7.44 (m, 1H), 7.22-7.28 (m, 2H),7.07-7.17 (m, 3H), 6.99 (t, J=8.0 Hz, 1H), 6.73 (d, J=16.8 Hz, 1H), 2.42(s, 3H).

Step 2:

(E/Z)-2-(3-bromostyryl)-1-chloro-3-methylbenzene was carbonylatedaccording to the method used in Example 50 except that additional DMFwas not added at the end of the reaction. Purification by flashchromatography (10 to 50% EtOAc-hexanes gradient) was conducted twice topartially separate the geometric isomers.(E)-3-(2-chloro-6-methylstyryl)benzaldehyde was isolated as an oil.Yield 0.0792 g, 22%), trans-/cis-isomer ratio 11.5:1. Trans-isomer: ¹HNMR (400 MHz, CDCl₃) δ 10.07 (s, 1H), 8.02 (t, J=2.0 Hz, 1H), 7.77-7.82(m, 2H), 7.55 (t, J=8.0 Hz, 1H), 7.28 (dd, J=9.2, 1.6 Hz, 1H), 7.24 (d,J=16.0 Hz, 1H), 7.15 (d, J=6.4 Hz, 1H), 7.11 (t, J=7.6 Hz, 1H), 6.86 (d,J=16.4 Hz, 1H), 2.44 (s, 3H).

Step 3:

(E)-3-(2-Chloro-6-methylstyryl)benzaldehyde was reacted withacetonitrile according to the procedure used in Example 50. Purificationby flash chromatography (10 to 70% EtOAc-hexanes gradient) gave(E)-3-(3-(2-chloro-6-methylstyryl)phenyl)-3-hydroxypropanenitrile as anoil. Yield 0.0708 g, 77%): ¹H NMR (400 MHz, CDCl₃) δ 7.51-7.53 (m, 2H),7.41 (t, J=8.0 Hz, 1H), 7.25-7.33 (m, 2H), 7.07-7.20 (m, 3H), 6.80 (d,J=16.8 Hz, 1H), 5.08 (t, J=6.4 Hz, 1H), 2.80 (dd, J=6.4, 0.8 Hz, 2H),2.43 (s, 3H).

Step 4:

(E)-3-(3-(2-Chloro-6-methylstyryl)phenyl)-3-hydroxypropanenitrile wasreduced according to the method used in Example 50 except that 3.3equivalents of LiAlH₄ were used in the reaction. Purification by flashchromatography (1:4:5 7 M NH₃ in MeOH: hexanes: EtOAc) was conductedtwice to give Example 82 as an oil. Yield (0.0230 g, 32%): ¹H NMR (400MHz, CDCl₃) δ 7.55 (s, 1H), 7.43 (d, J=7.6 Hz, 1H), 7.35 (t, J=7.6 Hz,1H), 7.25-7.29 (m, 2H), 7.07-7.20 (m, 3H), 6.80 (d, J=16.8 Hz, 1H), 5.00(dd, J=8.8, 3.2 Hz, 1H), 3.14 (ddd, J=12.4, 5.6, 4.0 Hz, 1H), 2.99 (ddd,J=13.2, 9.6, 4.0 Hz, 1H), 2.89 (br s, 2H), 2.43 (s, 3H), 1.88-1.94 (m,1H), 1.75-1.84 (m, 1H).

Example 83 Preparation of (E)-4-(3-(3-aminopropyl)styryl)heptan-4-ol

(E)-4-(3-(3-Aminopropyl)styryl)heptan-4-ol was prepared according to themethod used in Example 79.

Step 1:

4-Vinylheptan-4-ol was coupled to aryl bromide 104 and purified by flashchromatography (20 to 80% EtOAc-hexanes gradient) to afford(E)-2,2,2-trifluoro-N-(3-(3-(3-hydroxy-3-propylhex-1-enyl)phenyl)propyl)acetamideas an oil. Yield (0.4966 g, 81%): ¹H NMR (400 MHz, DMSO-d₃) δ 9.40 (brs, 1H), 7.19-7.21 (m, 3H), 7.02 (dt, J=6.0, 1.2 Hz, 1H), 6.44 (d, J=16.0Hz, 1H), 6.21 (d, J=16.0 Hz, 1H), 4.30 (s, 1H), 3.18 (q, J=6.4 Hz, 2H),2.56 (t, J=7.2 Hz, 2H), 1.78 (quint, J=7.4 Hz, 2H), 1.41-1.47 (m, 4H),1.18-1.36 (m, 4H), 0.83 (t, J=7.4 Hz, 6H).

Step 2:

(E)-2,2,2-Trifluoro-N-(3-(3-(3-hydroxy-3-propylhex-1-enyl)phenyl)propyl)acetamidewas deprotected according to the method used in Example 79 except that 3equivalents of K₂CO₃ were used. Following the reaction, solids wereremoved by filtration and the filtrate was concentrated under reducedpressure. The residue was dissolved in MeOH, dried over MgSO₄ andfiltered through Celite. After concentration under reduced pressure,purification by flash chromatography (7:3 EtOAc: hexanes to 7:2:1EtOAc:hexanes:7 M NH₃ in MeOH gradient) gave Example 83 as a singletrans isomer. Yield (0.348 g, quant.): ¹H NMR (400 MHz, DMSO-d₃) δ7.18-7.22 (m, 3H), 7.03-7.06 (m, 1H), 6.51 (d, J=16.0 Hz, 1H), 6.20 (d,J=16.4 Hz, 1H), 4.30 (br s, 1H), 2.66 (t, J=7.4 Hz, 2H), 2.64 (t, J=7.8Hz, 2H), 1.75-1.82 (m, 2H), 1.53-1.59 (m, 4H), 1.33-1.44 (m, 4H), 0.92(t, J=7.4 Hz, 6H).

Example 84 Preparation of (E)-1-(3-(3-aminopropyl)phenyl)hex-1-en-3-ol

(E)-1-(3-(3-Aminopropyl)phenyl)hex-1-en-3-ol was prepared according tothe method used in Example 79 with modifications.

Step 1:

Hex-1-en-3-ol was coupled to aryl bromide 104 according to the methodused in Example 79 except that 0.1 equivalents of tri(o-tolyl)phosphinewere used. Purification by flash chromatography (10 to 50% EtOAc-hexanesgradient) afforded(E)-2,2,2-trifluoro-N-(3-(3-(3-hydroxyhex-1-enyl)phenyl)propyl)acetamideas a yellow oil. Yield (0.258 g, 39%): ¹H NMR (400 MHz, CD₃OD) δ7.21-7.24 (m, 3H), 7.01-7.08 (m, 1H), 6.63 (dd, J=16.0, 0.8 Hz, 1H),6.21 (dd, J=16.0, 6.8 Hz, 1H), 4.19 (q, J=7.2 Hz, 1H), 2.63 (t, J=7.2Hz, 2H), 1.85-1.91 (m, 2H), 1.42-1.66 (m, 6H), 0.95 (t, J=7.6 Hz, 3H).

Step 2:

To a solution of(E)-2,2,2-trifluoro-N-(3-(3-(3-hydroxyhex-1-enyl)phenyl)propyl)acetamide(0.258 g, 0.78 mmol) in MeOH (10 mL) was added concentrated aqueousNH₄OH (10 mL). The reaction mixture was capped and stirred at roomtemperature overnight. The mixture was partitioned between diethyl etherand water and the combined organics were washed with brine, dried overNa₂SO₄ and concentrated under reduced pressure. Purification by flashchromatography (10 to 20% 7 M NH₃ in MeOH-EtOAc gradient) affordedExample 84 as a single trans isomer. Yield (0.110 g, 60%): ¹H NMR (400MHz, DMSO-d₆) δ 7.18-7.21 (m, 3H), 7.01-7.04 (m, 1H), 6.44 (dd, J=16.0,1.2 Hz, 1H), 6.21 (dd, J=16.0, 6.4 Hz, 1H), 4.07 (q, J=5.6 Hz, 1H),2.51-2.57 (m, 4H), 1.57-1.64 (m, 2H), 1.26-1.49 (m, 6H), 0.87 (t, J=7.2Hz, 3H).

Example 85 Preparation of (E)-4-(3-(2-aminoethoxy)styryl)heptan-4-ol

(E)-4-(3-(2-Aminoethoxy)styryl)heptan-4-ol was prepared according toScheme 36.

Step 1:

To a solution of 3-bromophenol (36.38 g, 210.3 mmol) in acetone (175 ml)was added K₂CO₃ (0.033 g, 237 mmol) and 2-bromoethanol (20 ml, 283.3mmol). The reaction mixture was heated at reflux under argon for 4 days.

After cooling to room temperature, the mixture was filtered and thefiltrate was concentrated under reduced pressure. The residue wasdissolved in diethyl ether (150 ml) and the solution was washedsuccessively with water, 10% aqueous NaOH, 5% aqueous NaOH, water, andbrine. The solution was dried over MgSO₄ and concentrated under reducedpressure to give 2-(3-bromophenoxyl)ethanol (106) as a light brown oil.Yield (21.07 g, 46%): ¹H NMR (400 MHz, CDCl₃) δ 7.14 (t, J=7.8 Hz, 1H),7.07-7.12 (m, 2H), 6.85 (ddd, J=7.8, 2.4, 1.3 Hz, 1H), 4.05-4.07 (m,2H), 3.93-3.97 (m, 2H), 2.11 (t, J=12.3 Hz. 1H).

Step 2:

To an ice cold mixture of 2-(3-bromophenoxyl)ethanol (106) (16.06 g,74.0 mmol) and triethylamine (9.12 g, 90.13 ml) in anhydrous CH₂Cl₂ (120ml) under argon was slowly added methane sulfonyl chloride (6 ml, 77.2mmol) The reaction mixture was stirred at 0° C. for 15 min. The mixturewas concentrated under reduced pressure and the residue was partitionedbetween EtOAc and water. The combined organics were washed with waterand brine, dried over MgSO₄, and concentrated under reduced pressure togive 2-(3-bromophenoxyl)ethyl methanesulfonate (107) as a light brownoil. This product was used in the next synthetic step without furtherpurification. Yield (21.32 g, 98%): ¹H NMR (400 MHz, CDCl₃) δ 7.16 (t,J=7.8 Hz, 1H), 7.11-7.14 (m, 1H), 7.06-7.07 (m, 1H), 6.39 (ddd, J=7.6,2.5, 1.8 Hz, 1H), 4.54-4.57 (m, 2H), 4.21-4.24 (m, 2H), 3.08 (s, 3H).

Step 3:

To a solution of mesylate 107 (24.05 g, 81.5 mmol) in anhydrous DMF (160ml) was added potassium phthalimide (15.53 g, 83.8 mmol) and thereaction mixture was stirred at 60° C. for 14 h. The mixture wasconcentrated under reduced pressure and the residue was partitionedbetween hexanes-EtOAc (7:1) and water. A precipitate formed which wascollected by filtration, washed excessively with water and hexanes, thendried under vacuum to give N-(2-(3-bromophenoxyl)ethyl)phthalimide (108)as white fluffy crystals (22.05 g). The second batch was collected byconcentrating the organic layer of the filtrate under reduced pressureand suspending the residue 10% EtOAc-hexanes. The mixture was washedwith water and the precipitate collected by filtration, washedexcessively with water, then hexanes and dried under vacuum to givephthalimide 108 (5.65 g). Combined yield (21.18 g, 98%). ¹H NMR (400MHz, CDCl₃) δ 7.87 (dd, J=5.2, 2.8 Hz, 2H), 7.73 (dd, J=5.6, 3.2 Hz,2H), 7.03-7.12 (m, 3H), 6.80 (ddd, J=8.0, 2.5, 1.4 Hz, 1H), 4.21 (t,J=6.9 Hz, 2H), 4.10 (t, J=6.0 Hz, 2H).

Step 4:

To a suspension of phthalimide 108 (22.82 g, 65.9 mmol) in absolute EtOH(200 ml) was added hydrazine hydrate (6 ml, 123.7 mmol) and the reactionmixture was heated at reflux under argon for 1.5 h. After cooling toroom temperature, the mixture was filtered and the filtrate wasconcentrated under reduced pressure. The residue was re-suspended inhexanes (100 ml) and the solids removed by filtration. The filtrate wasconcentrated under reduced pressure then the residue was taken up inEtOH and concentrated under reduced pressure. This procedure wasrepeated with toluene to give amine 109 as a thick yellow oil. Yield(10.63 g, 75%): ¹H NMR (400 MHz, CDCl₃) δ 7.11-7.15 (m, 1H), 7.06-7.09(m, 2H), 6.84 (ddd, J=8.0, 2.5, 1.2 Hz, 1H), 3.96 (t, J=5.3 Hz, 2H),3.07 (t, J=5.09 Hz, 2H), 1.43 (br s, 2H).

Step 5:

To a solution of amine 109 (10.63 g, 49.2 mmol) in anhydrous THF (80 ml)was added ethyl trifluoroacetate (12 ml, 100.6 mmol) and the reactionmixture was stirred at room temperature overnight. The resultingsolution was concentrated under reduced pressure and the residue wasdissolved in 50% EtOAc-hexanes. Purification by filtration through alayer of a silica gel, eluting with 50% EtOAc-hexanes gave bromide 110as a pale yellow oil which crystallized upon standing to a pale yellowsolid. Yield (13.69 g, 89%): ¹H NMR (400 MHz, CDCl₃) δ 7.16 (t, J=8.0Hz, 1H), 7.12-7.14 (m, 1H), 7.05-7.07 (m, 1H), 6.83 (ddd, J=7.6, 2.5,1.8 Hz, 1H), 6.75 (br s, 1H), 4.09 (t, J=4.9 Hz, 2H), 3.78 (q, J=5.5 Hz,2H).

Step 6:

4-Vinylheptan-4-ol was coupled to aryl bromide 110 according to themethod used in Example 84. Purification by flash chromatography (10 to40% EtOAc-hexanes gradient) gave trifluoroacetamide 111 as a light amberoil. Yield (0.417 g, 77%): ¹H NMR (400 MHz, CD₃OD) δ 7.19 (t, J=8.0 Hz,1H), 6.94-6.98 (m, 2H), 6.78 (ddd, J=8.4, 2.8, 0.8 Hz, 1H), 6.50 (d,J=16.4 Hz, 1H), 6.22 (d, J=16.0 Hz, 1H), 4.06-4.12 (m, 3H), 3.66 (t,J=5.6 Hz, 2H), 1.53-1.60 (m, 4H), 1.30-1.47 (m, 4H), 0.91 (t, J=7.2 Hz,6H).

Step 7:

Trifluoroacetamide 111 was deprotected according to the method used inExample 79 except that 5 equivalents of K₂CO₃ were used. The mixture wasconcentrated under reduced pressure and the residue was partitionedbetween EtOAc and water. The combined organics were washed with brine,dried over Na₂SO₄ and concentrated under reduced pressure to giveExample 85 as a single trans isomer. Yield (0.1134 g, 36%): ¹H NMR (400MHz, CD₃OD) δ 7.17-7.21 (m, 1H), 6.95-6.97 (m, 2H), 6.78-6.81 (m, 1H),6.50 (d, J=16.4 Hz, 1H), 6.22 (d, J=16.0 Hz, 1H), 4.00 (t, J=5.2 Hz,2H), 2.99 (t, J=5.2 Hz, 2H), 1.53-1.60 (m, 4H), 1.28-1.48 (m, 4H), 0.92(t, J=7.2 Hz, 6H).

Example 86 Preparation of(R,E)-3-amino-1-(3-(2,6-dichlorostyryl)phenyl)propan-1-ol

(R,E)-3-Amino-1-(3-(2,6-dichlorostyryl)phenyl)propan-1-ol is preparedaccording to Methods A, K and U and chiral reduction, as describedherein.

Example 87 Preparation of(S,E)-3-amino-1-(3-(2,6-dichlorostyryl)phenyl)propan-1-ol

(S,E)-3-Amino-1-(3-(2,6-dichlorostyryl)phenyl)propan-1-ol is preparedaccording to Methods A, K and U and chiral reduction, as describedherein.

Example 88 Preparation of(S,E)-3-(3-(2,6-dichlorostyryl)phenyl)-2-fluoropropan-1-amine

(S,E)-3-(3-(2,6-dichlorostyryl)phenyl)-2-fluoropropan-1-amine isprepared according to Methods A, R and X, as described herein.

Example 89 Preparation of(E)-3-(3-(2,6-dichlorostyryl)phenyl)-2,2-difluoropropan-1-amine

(E)-3-(3-(2,6-Dichlorostyryl)phenyl)-2,2-difluoropropan-1-amine wasprepared according to Scheme 37.

Step 1:

(E)-tert-butyl 3-(3-(2,6-dichlorostyryl)phenyl)-2-oxopropylcarbamate(112) (0.1633 g, 0.39 mmol) was stirred withbis(2-methoxyethyl)aminosulfur trifluoride (0.2 mL, 1.08 mmol) at roomtemperature for 1 h, 20 min. Additional bis(2-methoxyethyl)aminosulfurtrifluoride (0.2 mL, 1.08 mmol) was added and the mixture was stirred atroom temperature overnight. The mixture was purified by flashchromatography (10 to 40% EtOAc-hexanes gradient) to give difluoride 113as an oil. Yield (0.0694 g, 41%): ¹H NMR (400 MHz, CDCl₃) δ 7.49 (d,J=8.0 Hz, 1H), 7.43 (br s, 1H), 7.35 (t, J=7.6 Hz, 1H), 7.347 (d, J=8.0Hz, 2H), 7.23 (d, J=7.2 Hz, 1H), 7.09-7.14 (m, 3H), 4.82 (br s, 1H),3.54 (ddd, J=14.0, 14.0, 6.4 Hz, 2H), 3.21 (t, J=16.8 Hz, 2H), 1.46 (s,9H).

Step 2:

To a solution of difluoride 113 (0.0694 g, 0.16 mmol) in EtOAc (0.5 mL)was added a solution of HCl (3 mL of a 4.6 M solution in EtOAc, 13.8mmol) and the mixture was stirred for 3 h. The solid was collected byfiltration and dried under vacuum to give Example 89 hydrochloride as awhite solid. Yield (0.037 g, 62%): ¹H NMR (400 MHz, DMSO-d₆) δ 8.39 (brs, 3H), 7.59 (d, J=7.6 Hz, 1H), 7.52-7.54 (m, 3H), 7.41 (t, J=7.6 Hz,1H), 7.33 (t, J=8.0 Hz, 1H), 7.27 (d, J=7.6 Hz, 1H), 7.15 (d, J=16.4 Hz,1H), 7.08 (d, J=16.4 Hz, 1H), 3.36-3.45 (m, 4H).

Example 90 Preparation of(Z)-3-(3-(2-(2-methoxyethoxy)styryl)phenyl)-propan-1-amine

(Z)-3-(3-(2-(2-Methoxyethoxy)styryl)phenyl)-propan-1-amine was preparedaccording to the method used in Example 77.

Step 1:

Phthalimide 29 was coupled with 2-(2-methoxyethoxyl)benzylphosphoniumbromide according to the method used in Example 76 except that 2equivalents of phosphonium salt were used and the reaction was stirredfor 1 h at room temperature. Purification by flash chromatography (15%EtOAc-hexanes) gave(E/Z)-2-(3-(3-(2-(2-methoxyethoxy)styryl)phenyl)propyl)isoindoline-1,3-dioneas a brown oil. Yield (0.700 g, 58%).

Step 2:

(E/Z)-2-(3-(3-(2-(2-Methoxyethoxy)styryl)phenyl)propyl)isoindoline-1,3-dionewas deprotected following the method used in Example 76 except that thereaction was conducted at room temperature for 3 h. The mixture wasconcentrated under reduced pressure. The white solid was washed withdiethyl ether and the decanted solution was concentrated under reducedpressure. The residue was purified by Preparative HPLC (Method 2) togive Example 90 trifluoroacetate. Yield (0.030 g, 9%): ¹H NMR (DMSO-d₆)δ 7.66 (br s, 3H), 7.22 (t, J=8.0 Hz, 1H), 7.17 (t, J=8.0 Hz, 1H),7.03-7.12 (m, 5H), 6.76 (t, J=7.2 Hz, 1H), 6.64 (d, J=12.4 Hz, 1H), 6.59(d, J=12.0 Hz, 1H), 4.10 (t, J=4.4 Hz, 2H), 3.60 (t, J=4.8 Hz, 2H), 3.31(s, 3H), 2.73 (t, J=7.2 Hz, 2H), 1.72-1.77 (m, 2H), 1.17-1.90 (m, 2H).

Example 91 Preparation of(E)-3-(3-(3-methoxystyryl)phenyl)propan-1-amine

(E)-3-(3-(3-Methoxystyryl)phenyl)propan-1-amine was prepared accordingto the method used in Example 76.

Step 1:

Phthalimide 29 was coupled with (2-methoxybenzyl)triphenylphosphoniumbromide according to the method used in Example 76. Purification byflash chromatography (8% EtOAc-hexanes) gave(E)-2-(3-(3-(3-methoxystyryl)phenyl)propyl)isoindoline-1,3-dione a brownsemi-solid. Yield (0.090 g, 29%): ¹H NMR (400 MHz, CDCl₃) δ 7.81-7.83(m, 2H), 7.68-7.70 (m, 2H), 7.34 (s, 1H), 7.21-7.30 (m, 3H), 7.09-7.12(m, 2H), 7.05 (s, 3H), 6.82 (dd, J=8.0, 2.0 Hz, 1H), 3.86 (s, 3H), 3.78(t, J=7.2 Hz, 2H), 2.72 (t, J=7.6 Hz, 2H), 2.05-2.12 (m, 2H).

Step 2:

(E)-2-(3-(3-(3-Methoxystyryl)phenyl)propyl)isoindoline-1,3-dione wasdeprotected following the method used in Example 90 except that thereaction mixture was stirred overnight in methanol. Purification bypreparative thin layer chromatography on silica gel gave Example 97.Yield (0.010 g, 16%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.42 (s, 1H), 7.39 (d,J=7.6 Hz, 1H), 7.25-7.29 (m, 2H), 7.21 (d, J=4.0 Hz, 2H), 7.15-7.16 (m,2H), 7.09 (d, J=7.2 Hz, 1H), 6.82-6.83 (m, 1H), 3.77 (s, 3H), 2.53-2.63(m, 4H), 1.65-1.72 (m, 2H).

Example 92 Preparation of(E)-3-(3-(2-(1-methoxynaphthalen-2-yl)vinyl)phenyl)propan-1-amine

(E)-3-(3-(2-(1-Methoxynaphthalen-2-yl)vinyl)phenyl)propan-1-amine wasprepared according to the method used in Example 76.

Step 1:

(1-Methoxynaphthalen-2-ylmethyl)triphenylphosphonium bromide wasprepared according to the method used in Example 76, except that 1.1equivalents of triphenylphosphine were used in the reaction. Yield (1.25g, 70%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.84-7.88 (m, 5H), 7.67-7.70 (m,12H), 7.53-7.55 (m, 2H), 7.49 (d, J=8.4 Hz, 1H), 6.89 (d, J=8.4 Hz, 1H),5.15 (dd, J=15.6, 2.8 Hz, 2H), 3.74 (s, 3H).

Step 2:

Phthalimide 29 was coupled with(1-methoxynaphthalen-2-ylmethyl)triphenylphosphonium bromide accordingto the method used in Example 76 except that 2 equivalents of thephosphonium salt were used. Purification by flash chromatography (10%EtOAc-hexanes) gave(E/Z)-2-(3-(3-(2-(1-methoxynaphthalen-2-yl)vinyl)phenyl)propyl)isoindoline-1,3-dionea yellow oil. The mixture was further purified by Preparative HPLC togive the cis-isomer (0.034 g, 4% yield) as a colorless oil and thetrans-isomer (0.050 g, 7% yield) as a white solid. ¹H NMR (400 MHz,CDCl₃) δ 8.15 (d, J=8.4 Hz, 1H), 7.81-7.83 (m, 3H), 7.77 (d, J=8.8 Hz,1H), 7.68-7.71 (m, 2H), 7.63 (d, J=2.8 Hz, 1H), 7.59 (d, J=10.4 Hz, 1H),7.44-7.54 (m, 2H), 7.42 (s, 1H), 7.37 (d, J=12.0 Hz, 1H), 7.26 (t, J=7.6Hz, 1H), 7.17 (d, J=16.4 Hz, 1H), 7.12 (d, J=7.6 Hz, 1H), 3.99 (s, 3H),3.79 (t, J=7.2 Hz, 2H), 2.74 (t, J=7.6 Hz, 2H), 2.06-2.14 (m, 2H).

Step 3:

(E)-2-(3-(3-(2-(1-methoxynaphthalen-2-yl)vinyl)phenyl)propyl)isoindoline-1,3-dionewas deprotected following the method used in Example 76 except that thereaction was conducted in MeOH at room temperature for 2 h. The reactionwas concentrated under reduced pressure. The white solid was washed withdiethyl ether and the decanted solution was concentrated under reducedpressure. The residue was purified by preperative thin layerchromatography on silica gel (1:10:89 NH₄OH:MeOH:CH₂Cl₂) to give Example92. Yield (0.020 g, 57%): ¹H NMR (400 MHz, DMSO-d₆) δ 8.07 (d, J=8.4 Hz,1H), 7.91 (d, J=8.8 Hz, 2H), 7.71 (d, J=8.8 Hz, 1H), 7.46-7.57 (m, 5H),7.38 (d, J=16.4 Hz, 1H), 7.31 (t, J=7.6 Hz, 1H), 7.13 (d, J=7.6 Hz, 1H),3.91 (s, 3H), 2.62-2.66 (m, 2H), 2.57 (t, J=6.8 Hz, 2H), 1.64-1.72 (m,2H).

Example 93 Preparation of (Z)-3-(3-(4-chlorostyryl)phenyl)propan-1-amine

(Z)-3-(3-(4-Chlorostyryl)phenyl)propan-1-amine was prepared according tothe method used in Example 76.

Step 1:

Phthalimide 29 was coupled with (4-chlorobenzyl)triphenylphosphoniumbromide according to the method used in Example 76. Purification byflash chromatography (7% EtOAc-hexanes) gave(E)-2-(3-(3-(4-chlorostyryl)phenyl)propyl)isoindoline-1,3-dione (0.120g, 10%) and(Z)-2-(3-(3-(4-chlorostyryl)phenyl)propyl)isoindoline-1,3-dione (0.150g, 13%) as yellow solids. Cis-isomer: ¹H NMR (400 MHz, CDCl₃) δ7.79-7.86 (m, 2H), 7.68-7.73 (m, 3H), 7.12-7.17 (m, 5H), 7.00-7.05 (m,3H), 6.58 (d, J=12.0 Hz, 1H), 6.50 (d, J=12.0 Hz, 1H), 3.68 (t, J=7.2Hz, 2H), 2.60 (t, J=7.6 Hz, 2H), 1.90-1.97 (m, 2H).

Step 2:

From another preparation, a mixture of(E/Z)-2-(3-(3-(4-chlorostyryl)phenyl)propyl)isoindoline-1,3-dione wasdeprotected following the method used in Example 76. The crude productwas purified by Preparative HPLC (Method 2) to give Example 93trifluoroacetate. Yield (0.020 g, 16%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.72(br s, 3H), 7.31-7.34 (m, 2H), 7.21-7.25 (m, 3H), 7.09-7.10 (m, 2H),7.05 (d, J=7.6 Hz, 1H), 6.67 (d, J=12.4 Hz, 1H), 6.60 (d, J=12.0 Hz,1H), 2.75 (t, J=7.2 Hz, 2H), 2.56 (t, J=7.6 Hz, 2H), 1.73-1.81 (m, 2H).

Example 94 Preparation of(E)-3-(3-(2-(biphenyl-2-yl)vinyl)phenyl)propan-1-amine

(E)-3-(3-(2-(Biphenyl-2-yl)vinyl)phenyl)propan-1-amine was preparedaccording to the method used in Example 76.

Step 1:

Phthalimide 29 was coupled with biphenyl-2-ylmethyl triphenylphosphoniumbromide. Purification by flash chromatography (5% EtOAc-hexanes) gave(E)-2-(3-(3-(2-(biphenyl-2-yl)vinyl)phenyl)propyl)isoindoline-1,3-dione(0.100 g, 13%) and(E/Z)-2-(3-(3-(2-(biphenyl-2-yl)vinyl)phenyl)propyl)isoindoline-1,3-dione(0.230 g, 30%) as colorless oils. Trans-isomer: ¹H NMR (400 MHz, CDCl₃)δ 781-7.83 (m, 2H), 7.72-7.74 (m, 1H), 7.68-7.70 (m, 2H), 7.63-7.65 (m,2H), 7.42-7.52 (m, 6H), 7.35-7.40 (m, 2H), 7.22-7.24 (m, 1H), 7.09-7.14(m, 3H), 3.79 (t, J=7.2 Hz, 2H), 2.73 (t, J=7.6 Hz, 2H), 2.05-2.13 (m,2H).

Step 2:

(E)-2-(3-(3-(2-(biphenyl-2-yl)vinyl)phenyl)propyl)isoindoline-1,3-dionewas deprotected following the method used in Example 76 except that thereaction was conducted at room temperature for 36 h. Purification bypreperative thin layer chromatography on silica gel gave Example 94.Yield (0.051 g, 71%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.84 (d, J=7.2 Hz,1H), 7.71 (br s, 3H), 7.49 (t, J=7.2 Hz, 1H), 7.33-7.44 (m, 7H),7.16-7.30 (m, 4H), 7.10 (d, J=7.2 Hz, 1H), 7.03 (d, J=16.4 Hz, 1H), 2.78(t, J=7.6 Hz, 2H), 2.62 (t, J=7.6 Hz, 2H), 1.78-1.85 (m, 2H).

Example 95 Preparation of(Z)-3-(3-(2-(naphthalen-1-yl)vinyl)phenyl)propan-1-amine

(Z)-3-(3-(2-(Naphthalen-1-yl)vinyl)phenyl)propan-1-amine was preparedaccording to the method used in Example 76.

Step 1:

Phthalimide 29 was coupled with (naphthalen-1-ylmethyl)triphenylphosphonium bromide according to the method used inExample 76. Purification by flash chromatography (10% EtOAc-hexanes)gave(E/Z)-2-(3-(3-(2-(naphthalen-1-yl)vinyl)phenyl)propyl)isoindoline-1,3-dionea brown oil. Yield (0.090 g, 46%).

Step 2:

(E/Z)-2-(3-(3-(2-(naphthalen-1-yl)vinyl)phenyl)propyl)isoindoline-1,3-dionewas deprotected following the method used in Example 76. The reactionwas concentrated under reduced pressure. The white solid was washed withdiethyl ether and the decanted solution was concentrated under reducedpressure. The residue was purified by Preparative HPLC (Method 2) togive Example 92 trifluoroacetate. Yield (0.020 g, 14%): ¹H NMR (400 MHz,DMSO-d₆) δ 7.96-8.02 (m, 2H), 7.88 (d, J=8.0 Hz, 1H), 7.51-7.57 (m, 5H),7.42 (t, J=7.6 Hz, 1H), 7.32 (d, J=6.8 Hz, 1H), 7.12 (d, J=12.4 Hz, 1H),7.03 (t, J=7.2 Hz, 1H), 6.94-6.97 (m, 2H), 6.87 (d, J=12.4 Hz, 1H), 6.83(d, J=7.6 Hz, 1H), 2.64 (t, J=7.6 Hz, 2H), 2.43 (t, J=7.6 Hz, 2H),1.60-1.68 (m, 2H).

Example 96 Preparation of(Z)-3-(3-(2-(3-methoxynaphthalen-2-yl)vinyl)phenyl)propan-1-amine

(Z)-3-(3-(2-(3-Methoxynaphthalen-2-yl)vinyl)phenyl)propan-1-amine wasprepared according to the method used in Example 77.

Step 1:

Phthalimide 29 was coupled with(3-methoxynaphthalen-2-ylmethyl)triphenylphosphonium bromide accordingto the method used in Example 76. Purification by flash chromatography(17% EtOAc-hexanes) gave(E/Z)-2-(3-(3-(2-(3-methoxynaphthalen-2-yl)vinyl)phenyl)propyl)isoindoline-1,3-dionea yellow oil. Yield (0.500 g, 40%).

Step 2:

(E/Z)-2-(3-(3-(2-(3-methoxynaphthalen-2-yl)vinyl)phenyl)propyl)isoindoline-1,3-dionewas deprotected following the method used in Example 76. The crudeproduct was purified by Preparative HPLC (Method 2) to give Example 96trifluoroacetate. Yield (0.030 g, 10%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.81(d, J=8.0 Hz, 1H), 7.69 (br s, 3H), 7.60 (s, 1H), 7.55 (d, J=8.0 Hz,1H), 7.42 (ddd, J=9.2, 6.8, 1.2 Hz, 1H), 7.38 (s, 1H), 7.27 (ddd, J=8.4,7.2, 1.2 Hz, 1H), 7.08-7.12 (m, 2H), 7.00-7.03 (m, 2H), 6.74 (d, J=12.4Hz, 1H), 6.70 (d, J=12.4 Hz, 1H), 3.88 (s, 3H), 2.65-2.73 (m, 2H),2.48-2.50 (m, 2H), 1.67-1.75 (m, 2H).

Example 97 Preparation of (E)-3-(3-(3-chlorostyryl)phenyl)propan-1-amine

(E)-3-(3-(3-Chlorostyryl)phenyl)propan-1-amine was prepared according tothe method used in Example 76.

Step 1:

Phthalimide 29 was coupled with 3-chlorobenzyltriphenylphosphoniumbromide. Purification by flash chromatography (5% EtOAc-hexanes) gave(E)-2-(3-(3-(3-chlorostyryl)phenyl)propyl)isoindoline-1,3-dione (0.100g, 30%) and(Z)-2-(3-(3-(3-chlorostyryl)phenyl)propyl)isoindoline-1,3-dione (0.180g, 30%) as colorless oils. Trans-isomer: ¹H NMR (400 MHz, CDCl₃) δ7.80-7.84 (m, 2H), 7.68-7.72 (m, 2H), 7.50 (t, J=1.6 Hz, 1H), 7.37 (dt,J=7.6, 1.2 Hz, 1H), 7.35 (s, 1H), 7.29 (d, J=7.6 Hz, 1H), 7.21-7.26 (m,3H), 7.12 (dt, J=6.4, 2.0 Hz, 1H), 7.03 (d, J=4.4 Hz, 2H), 3.78 (t,J=7.2 Hz, 2H), 2.72 (t, J=7.6 Hz, 2H), 2.05-2.12 (m, 2H).

Step 2:

From another preparation, a mixture of(E/Z)-2-(3-(3-(3-chlorostyryl)phenyl)propyl)isoindoline-1,3-dione wasdeprotected. The crude product was purified by Preparative HPLC (Method2) to give Example 97 trifluoroacetate as a white solid. Yield (0.040 g,67%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.73 (br s, 3H), 7.70 (s, 1H), 7.55(d, J=7.6 Hz, 1H), 7.44-7.48 (m, 2H), 7.41 (t, J=8.0 Hz, 1H), 7.36 (d,J=4.0 Hz, 1H), 7.32-7.35 (m, 2H), 7.24 (t, J=16.4 Hz, 1H), 7.15 (d,J=7.6 Hz, 1H), 2.81 (t, J=7.2 Hz, 2H), 2.67 (t, J=7.6 Hz, 2H), 1.83-1.91(m, 2H).

Example 98 Preparation of (E)-3-(3-(2-butoxystyryl)phenyl)propan-1-amine

(E)-3-(3-(2-butoxystyryl)phenyl)propan-1-amine is prepared according tothe method used in Example 32.

Example 99 Preparation of(E)-3-(3-(4-methoxystyryl)phenyl)propan-1-amine

(E)-3-(3-(4-Methoxystyryl)phenyl)propan-1-amin is prepared according tothe method used in Example 32.

Example 100 Preparation of(E)-1-(3-(3-aminopropyl)phenyl)-3-ethylpent-1-en-3-ol

(E)-1-(3-(3-aminopropyl)phenyl)-3-ethylpent-1-en-3-ol was preparedaccording to the method used in Example 79.

Step 1:

3-Ethylpent-1-en-3-ol was coupled to aryl bromide 104. Purification byflash chromatography (20 to 65% EtOAc-hexanes gradient) gave(E)-N-(3-(3-(3-ethyl-3-hydroxyl)ent-1-enyl)phenyl)propyl)-2,2,2-trifluoroacetamideas a light yellow syrup. Yield (0.401 g, 90%): ¹H NMR (400 MHz, DMSO-d₃)δ 9.41 (br s, 1H), 7.19-7.22 (m, 3H), 7.01-7.04 (m, 1H), 6.46 (d, J=16.4Hz, 1H), 6.18 (d, J=16.4 Hz, 1H), 4.27 (s, 1H), 3.18 (q, J=6.8 Hz, 2H),2.55 (t, J=7.6 Hz, 2H), 1.76-1.79 (m, 2H), 1.49 (q, J=7.6 Hz, 4H), 0.79(t, J=7.6 Hz, 6H).

Step 2:

(E)-N-(3-(3-(3-Ethyl-3-hydroxyl)ent-1-enyl)phenyl)propyl)-2,2,2-trifluoroacetamidewas deprotected then purified by flash chromatography (80 to 100%EtOAc-hexanes then 8-10% 7 M NH₃ in MeOH-EtOAc gradient) to give Example100 as a colorless oil. Yield (0.2374 g, 85%): ¹H NMR (400 MHz, DMSO-d₆)7.17-7.20 (m, 3H), 7.00-7.02 (m, 1H), 6.45 (d, J=16.0 Hz, 1H), 6.17 (d,J=16.0 Hz, 1H), 4.27 (s, 1H), 2.50-2.57 (m, 4H), 1.57-1.64 (m, 2H), 1.49(q, J=7.6 Hz, 4H), 1.36 (br s, 2H), 0.79 (t, J=7.6 Hz, 6H).

Example 101 Preparation of (E)-3-(3-(3-aminopropyl)phenyl)prop-2-en-1-ol

(E)-3-(3-(3-Aminopropyl)phenyl)prop-2-en-1-ol is prepared according tothe method used in Example 79.

Example 102 Preparation of(E)-3-(3-(3-methoxyprop-1-enyl)phenyl)propan-1-amine-ol

(E)-3-(3-(3-Methoxyprop-1-enyl)phenyl)propan-1-amine-ol was preparedaccording to the method used in Example 79.

Step 1:

Allyl methyl ether was coupled to aryl bromide 104 according to themethod used in Example 84. Purification by flash chromatography twice(20 to 40% EtOAc-hexanes gradient) gave(E)-2,2,2-trifluoro-N-(3-(3-(3-methoxyprop-1-enyl)phenyl)propyl)acetamideas a yellow oil. Yield (0.060 g, 6%): ¹H NMR (400 MHz, CDCl₃) δ7.20-7.30 (m, 3H), 7.05-7.12 (m, 1H), 6.59 (d, J=16.0 Hz, 1H), 6.22-6.32(m, 2H), 3.36-3.40 (m, 5H), 2.64-2.72 (m, 2H), 1.88-2.0 (m, 4H).

Step 2:

(E)-2,2,2-Trifluoro-N-(3-(3-(3-methoxyprop-1-enyl)phenyl)propyl)acetamidewas deprotected according to the method used in Example 84. Purificationby flash chromatography (0 to 10% 7 M NH₃ in MeOH-EtOAc) gave Example102 as a colorless oil. Yield (0.023 g, 56%): ¹H NMR (400 MHz, CDCl₃) δ7.21-7.22 (m, 3H), 7.05-7.10 (m, 1H), 6.58 (dt, J=16.0, 1.2 Hz, 1H),6.26 (dt, J=16.0, 6.0 Hz, 1H), 4.08 (dd, J=6.0, 1.6 Hz, 2H), 3.36 (s,3H), 2.72 (t, J=6.8 Hz, 2H), 2.61 (t, J=8.0 Hz, 2H), 1.71-1.80 (m, 2H),1.18-1.28 (m, 2H).

Example 103 Preparation of(E)-1-(3-(3-aminopropyl)phenyl)-3-methylhex-1-en-3-ol

(E)-1-(3-(3-Aminopropyl)phenyl)-3-methylhex-1-en-3-ol is preparedaccording to the method used in Example 79.

Example 104 Preparation of(E)-1-(3-(3-aminopropyl)phenyl)-3-ethylhex-1-en-3-ol

(E)-1-(3-(3-Aminopropyl)phenyl)-3-ethylhex-1-en-3-ol is preparedaccording to the method used in Example 79.

Example 105 Preparation of(R,E)-3-amino-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-ol

(R,E)-3-Amino-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-ol(R/S stereochemistry is assumed not assigned) was prepared according toScheme 38.

Step 1:

To a solution of(E)-3-amino-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-ol(Example 25) (0.5137 g, 1.57 mmol) in CH₂Cl₂ (8 mL) was addeddiisopropyl ethylamine (0.33 mL, 1.90 mmol) and a solution of9-fluorenylmethoxycarbonyl chloride (0.4875 g, 1.88 mmol) in CH₂Cl₂ (2mL). The reaction mixture was stirred for 35 min then concentrated underreduced pressure. Purification by flash chromatography (10 to 70%EtOAc-hexanes gradient) gave alcohol 114 as an oil. Yield (0.5564 g,68%).

Step 2:

To a solution of alcohol 114 (0.5564, 1.07 mmol) in CH₂Cl₂ (20 mL) wasadded MnO₂ (3.10 g, 35.7 mmol) and the mixture was stirred at roomtemperature overnight. Solids were removed from the mixture byfiltration through a pad of silica gel and the filtrate was concentratedunder reduced pressure. Purification by flash chromatography (10 to 70%EtOAc-hexanes gradient) gave ketone 115 as an oil. Yield (0.4391 g,79%): ¹H NMR (400 MHz, CDCl₃) δ 7.98 (br s, 1H), 7.79 (d, J=8.0 Hz, 1H),7.73 (d, J=7.6 Hz, 2H), 7.62 (d, J=7.6 Hz, 1H), 7.57 (d, J=7.6 Hz, 2H),7.42 (t, J=7.6 Hz, 1H), 7.39 (t, J=7.6 Hz, 2H), 7.28 (t, J=7.6 Hz, 1H),6.77 (dd, J=16.0, 0.8 Hz, 1H), 6.39 (dd, J=16.0, 0.8 Hz, 1H), 5.43 (t,J=6.0 Hz, 1H), 4.38 (d, J=6.8 Hz, 2H), 4.19 (t, J=6.8 Hz, 1H), 3.65 (q,J=5.6 Hz, 2H), 3.25 (t, J=5.2 Hz, 2H), 2.06 (t, J=6.0 Hz, 2H), 1.77 (s,3H), 1.63-1.69 (m, 2H), 1.49-1.52 (m, 2H), 1.08 (s, 6H).

Step 3:

Preparation of (−)-B-chlorodiisopinocampheylborane solution((−)-DIP-Cl): To an ice-cold solution of (−)-α-pinene (1.0042 g, 7.4mmol) in hexanes (6 mL) under argon was added chloroborane-methylsulfide complex (0.4 mL, 3.84 mmol) slowly. Additional (−)-α-pinene(0.17 mL, 1.09 mmol) was added and the mixture was stirred for 5 minthen allowed to warm to room temperature over 3 min. The resultingsolution was approximately 0.5 M.

To a −25° C. solution of ketone 115 (0.2117 g, 0.41 mmol) anddiisopropyl ethylamine (0.020 mL, 0.115 mmol) in THF (2.5 mL) was addeda solution of (−)-DIP-Cl (1.6 mL of the 0.5 M solution, 0.80 mmol) over5 min. The reaction mixture was cooled to −78° C. for 5 min then allowedto warm to room temperature. After stirring 30 min, additional(−)-DIP-Cl (1.6 mL of 0.5 M solution, 0.80 mmol) was added and themixture was stirred for 2 h. Acetone (5 mL) was added then the mixturewas concentrated under reduced pressure. The residue was partitionedbetween EtOAc and brine and the combined organics were dried over MgSO₄and concentrated under reduced pressure. Purification by flashchromatography (10 to 50% EtOAc-hexanes gradient) gave alcohol 116 as anoil. Yield (0.1155 g, 54%): ¹H NMR (400 MHz, CDCl₃) δ 7.76 (d, J=7.6 Hz,2H), 7.60 (d, J=7.2 Hz, 2H), 7.27-7.42 (m, 7H), 7.19 (d, J=6.4 Hz, 1H),6.70 (dd, J=16.0, 0.8 Hz, 1H), 6.34 (d, J=16.4 Hz, 1H), 5.14 (br s, 1H),4.72 (t, J=7.2 Hz, 1H), 4.40-4.50 (m, 2H), 4.22 (t, J=6.8 Hz, 1H), 3.70(t, J=6.4 Hz, 0.5H), 3.59 (t, J=6.8 Hz, 0.5H), 3.53-3.59 (m, 1H),3.26-3.31 (m, 1H), 2.02-2.05 (m, 2H), 1.85-1.93 (m, 2H), 1.75 (s, 3H),1.61-1.67 (m, 2H), 1.48-1.51 (m, 1H), 1.06 (s, 6H).

Step 4:

To a solution of alcohol 116 (0.0.0608 g, 0.117 mmol) in THF (2 mL) wasadded 1,8-diazabicyclo[5.4.0]undec-7-ene (0.034 g, 0.22 mmol). Themixture was stirred at room temperature for 20 min then concentratedunder reduced pressure. The residue was partitioned between EtOAc andbrine and the combined organics were dried over Na₂SO₄ and concentratedunder reduced pressure. Purification by flash chromatography (5:9:1 to5:5:1 EtOAc/hexanes/7 M NH₃ in MeOH gradient) gave Example 105 as anoil. Yield (0.011 g, 32%): The ¹H NMR data was consistent with that ofExample 25. Chiral HPLC: 92.9% major enantiomer (AUC), t_(R)=18.042 min(minor enantiomer: 7.1%, t_(R)=20.413 min).

Example 106 Preparation of(S,E)-3-amino-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-ol

(S,E)-3-Amino-1-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propan-1-ol(R/S stereochemistry is assumed, not assigned) was prepared according tothe method used in Example 105 with modifications.

Step 1:

Ketone 115 was reduced with a freshly prepared solution of(+)-B-chlorodiisopinocampheylborane solution ((+)-DIP-Cl) according tothe method used in Example 105 to give (S,E)-(9H-fluoren-9-yl)methyl3-hydroxy-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propylcarbamateas an oil. Yield (0.1096 g, 51%).

Step 2:

(S,E)-(9H-fluoren-9-yl)methyl3-hydroxy-3-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)propylcarbamatewas deprotected and purified as described in Example 105 to give Example106 as an oil. Yield (0.0140 g, 39%). The ¹H NMR data was consistentwith that of Example 25. Chiral HPLC; 96.0% major enantiomer (AUC),t_(R)=20.282 min (minor enantiomer: 3.9%, t_(R)=20.282 min).

Example 107 Preparation of(E)-3-(5-(2-chloro-6-(methylthio)styryl)-2-methoxyphenyl)propan-1-amine

(E)-3-(5-(2-Chloro-6-(methylthio)styryl)-2-methoxyphenyl)propan-1-aminewas prepared according to Scheme 39.

Step 1:

To a solution of dichloroarene 102 (0.3772 g, 0.87 mmol) in DMF (9 mL)was added sodium thiomethoxide (0.1176, 1.68 mmol). The mixture washeated at 100° C. for 2 h then cooled to room temperature andconcentrated under reduced pressure. The residue was partitioned betweenEtOAc and water and the combined organics were washed with brine, driedover MgSO₄ and concentrated under reduced pressure. The reaction andextractive workup were repeated and the combined products were purifiedby flash chromatography (10 to 100% EtOAc-hexanes gradient) to givesulphide 126 as a white solid. Yield (0.3005 g, 82%): ¹H NMR (400 MHz,CDCl₃) δ 7.39 (dd, J=8.4, 2.4 Hz, 1H), 7.33 (d, J=2.4 Hz, 1H), 7.21 (dd,J=8.4, 2.4 Hz, 1H), 7.15 (t, J=8.0 Hz, 1H), 7.11 (dd, J=7.6, 1.6 Hz,1H), 6.94 (d, J=8.4 Hz, 2H), 6.89 (d, J=8.4 Hz, 1H), 6.77 (br s, 1H),3.87 (s, 3H), 3.23 (q, J=6.4 Hz, 2H), 2.73 (t, J=7.6 Hz, 2H), 2.45 (s,3H), 1.87-1.94 (m, 2H).

Step 2:

To a solution of sulphide 126 (0.1127 g, 0.25 mmol) in 7 M NH₃ in MeOH(8 mL) was added 25% aqueous NH₄OH (2 mL). The mixture was stirred atroom temperature overnight then concentrated under reduced pressure.Purification by flash chromatography (5:5:1 EtOAc:hexanes:7 M NH₃ inMeOH) gave Example 107 as an oil. Yield (0.0680 g, 77%): ¹H NMR (400MHz, CDCl₃) δ 7.34-7.36 (m, 2H), 7.20 (dd, J=8.4, 2.4 Hz, 1H), 7.13 (t,J=7.6 Hz, 1H), 7.10 (dd, J=8.0, 1.6 Hz, 1H), 6.97 (d, J=16.4 Hz, 1H),6.92 (d, J=16.8 Hz, 1H), 6.83 (dd, J=7.2, 0.8 Hz, 1H), 3.84 (s, 3H),2.73 (t, J=6.8 Hz, 2H), 2.68 (t, J=8.0 Hz, 2H), 2.41 (s, 3H), 1.74-1.78(m, 2H), 1.18 (br s, 2H).

Example 108 Preparation of(E/Z)-3-(3-(2-(2-methoxynaphthalen-1-yl)vinyl)phenyl)propan-1-amine

(E/Z)-3-(3-(2-(2-Methoxynaphthalen-1-yl)vinyl)phenyl)propan-1-amine wasprepared according to the method used in Example 76.

Step 1:

Phthalimide 29 was coupled with(2-methoxynapthalen-1-ylmethyl)triphenylphosphonium bromide according tothe method used in Example 76 except that 2 equivalents of thephosphonium bromide were used. Purified by flash chromatography (10%EtOAc-hexanes) gave(E/Z)-2-(3-(3-(2-(2-methoxynaphthalen-1-yl)vinyl)phenyl)propyl)isoindoline-1,3-dioneas an oil. Yield (0.160 g, 35%). Trans-isomer: ¹H NMR (400 MHz, DMSO-d₆)δ 8.24 (d, J=8.8 Hz, 1H), 7.88-7.92 (m, 2H), 7.81-7.86 (m, 4H), 7.58 (d,J=16.4 Hz, 1H), 7.49-7.53 (m, 3H), 7.37-7.43 (m, 2H), 7.28 (t, J=7.6 Hz,1H), 7.14-7.16 (m, 1H), 7.13 (d, J=16.8 Hz 1H), 3.97 (s, 3H), 3.65 (t,J=7.2 Hz, 2H), 2.69 (t, J=6.8 Hz, 2H), 1.94-2.00 (m, 2H).

Step 2:

(E/Z)-2-(3-(3-(2-(2-methoxynaphthalen-1-yl)vinyl)phenyl)propyl)isoindoline-1,3-dionewas deprotected following the method used in Example 92. The residue waspurified by Preparative HPLC (Method 2) to give Example 108trifluoroacetate. Yield (0.080 g, 51%): trans-/cis-isomer ratio 2:1.Trans-isomer: ¹H NMR (400 MHz, DMSO-d₆) δ 8.21 (d, J=8.4 Hz, 1H), 7.91(t, J=8.8 Hz, 2H), 7.73 (br s, 3H), 7.59 (d, J=16.4 Hz, 1H), 7.50-7.52(m, 4H), 7.33-7.45 (m, 2H), 7.12-7.45 (m, 2H), 3.98 (s, 3H), 2.75-2.86(m, 2H), 2.70 (t, J=7.6 Hz, 2H), 1.86-1.94 (m, 2H).

Example 109 Preparation of(Z)-3-(3-(2-propoxystyryl)phenyl)propan-1-amine

(Z)-3-(3-(2-Propoxystyryl)phenyl)propan-1-amine was prepared accordingto the method used in Example 76.

Step 1:

2-Propoxybenzyltriphenylphosphonium bromide was prepared from2-propoxylbenzyl bromide to give a white solid. Yield (2.4 g, 37%): ¹HNMR (400 MHz, DMSO-d₆) δ 7.87-7.92 (m, 3H), 7.72 (dt, J=8.0, 3.2 Hz,6H), 7.57-7.63 (m, 6H), 7.22-7.32 (m, 1H), 7.00-7.03 (m, 1H), 6.85 (d,J=8.4 Hz, 1H), 6.81 (t, J=7.2 Hz, 1H), 4.93 (d, J=14.4 Hz, 2H), 3.39 (t,J=6.8 Hz, 2H), 1.26-1.35 (m, 2H), 0.77 (t, J=7.2 Hz, 3H).

Step 2:

Phthalimide 29 was coupled with 2-propoxybenzyltriphenylphosphoniumbromide according to the method used in Example 76 except that thereaction was stirred for 1 h at room temperature after the addition ofpotassium tert-butoxide. Purification by flash chromatography (15%EtOAc-hexanes) gave(E/Z)-2-(3-(3-(2-propoxystyryl)phenyl)propyl)isoindoline-1,3-dione ayellow oil. Yield (0.080 g, 31%); trans-/cis-isomer ratio 1.3:1.Trans-isomer: ¹H NMR (400 MHz, CDCl₃) δ 7.80-7.85 (m, 2H), 7.67-7.72 (m,2H), 7.56-7.58 (m, 1H), 7.44 (d, J=16.4 Hz, 1H), 7.31 (s, 1H), 6.93-7.24(m, 6H), 6.89 (d, J=8.4 Hz, 1H), 4.00 (t, J=6.4 Hz, 2H), 3.78 (t, J=7.2Hz, 2H), 2.71 (t, J=7.6 Hz, 2H), 1.93-2.11 (m, 2H), 1.86-1.94 (m, 2H),1.10 (t, J=7.2 Hz, 3H).

Step 3:

(E/Z)-2-(3-(3-(2-propoxystyryl)phenyl)propyl)isoindoline-1,3-dione wasdeprotected following the method used in Example 76 except that thereaction was heated at 70° C. for 1.5 h. After cooling to roomtemperature, the mixture was concentrated under reduced pressure. Thewhite solid was washed with diethyl ether and the decanted solution wasconcentrated under reduced pressure. The residue was purified byPreparative HPLC (Method 2) to give Example 109 trifluoroacetate. Yield(0.120 g, 38%): ¹H NMR (400 MHz, CDCl₃) δ 7.84 (br s, 3H), 7.09-7.18 (m,4H), 6.99 (s, 1H), 6.92-6.94 (m, 1H), 6.86 (d, J=8.4 Hz, 1H), 6.68-6.72(m, 2H), 6.55 (d, J=12.4 Hz, 1H), 3.92 (t, J=6.4 Hz, 2H), 2.80 (t, J=7.6Hz, 2H), 2.53 (t, J=7.2 hz, 2H), 1.60-1.89 (m, 7H), 1.01 (t, J=7.2 Hz,3H).

Example 110 Preparation of(E)-3-(3-(2-phenylprop-1-enyl)phenyl)propan-1-amine

(E)-3-(3-(2-Phenylprop-1-enyl)phenyl)propan-1-amine was preparedaccording to the method shown in Scheme 40.

Step 1:

To a cooled solution (−78° C.) of diethyl 1-phenylethylphosphonate(1.016 g, 4.20 mmol) in anhydrous THF (10 mL) was added a solution ofn-BuLi (2.5M in hexanes, 1.5 mL, 3.75 mmol) and the mixture was stirredat −78° C. for 10 min under argon. A solution of aldehyde 29 (0.485 g,1.65 mmol) in anhydrous THF (10 mL) was added to the reaction mixtureand the mixture was stirred at −78° C. for 10 min, then allowed to warmto room temperature over 1 h. Saturated aqueous NH₄Cl was added to themixture followed by EtOAc. The layers were separated and the aqueouslayer was extracted with EtOAc. The combined organic layers were washedwith brine, dried with anhydrous MgSO₄, filtered and concentrated underreduced pressure. Purification by flash chromatography (10 to 30%EtOAc/hexane gradient) gave 118 as colorless oil. Yield (0.092 g, 15%):¹H NMR (400 MHz, DMSO-d₆) δ 7.78-7.84 (m, 4H), 7.51-7.55 (m, 2H),7.33-7.38 (m, 2H), 7.20-7.29 (m, 3H), 7.06-7.16 (m, 2H), 6.80 (s, 1H),3.60 (t, J=7.0 Hz, 2H), 2.63 (t, J=7.2 Hz, 2H), 2.19 (d, J=1.4 Hz, 3H),1.88-1.96 (m, 2H).

Step 2:

Deprotection of(E)-2-(3-(3-(2-phenylprop-1-enyl)phenyl)propyl)isoindoline-1,3-dione(118) with hydrazine following the method described in Example 18 exceptthat EtOH was used as the solvent and the reaction mixture was stirredat room temperature for 16 hrs. Purification by flash chromatography (20to 100% 20% 7N NH₃/MeOH in EtOAc/hexanes gave Example 110 as a colorlessoil. Yield (0.018 g, 30%). ¹H NMR (400 MHz, CD₃OD) δ 7.48-7.52 (m, 2H),7.30-7.35 (m, 2H), 7.20-7.28 (m, 2H), 7.14-7.19 (m, 2H), 7.05-7.09 (m,1H), 6.80 (s, 1H), 2.62-2.68 (m, 4H), 2.23 (d, J=1.4 Hz, 3H), 1.74-1.82(m, 2H). ¹³C NMR (100 MHz, CD₃OD) δ 144.1, 142.1, 138.6, 137.3, 129.1,128.2, 128.0, 127.6, 127.0, 126.52, 126.49, 125.8, 40.9, 33.1, 16.6. ESIMS m/z 252.3 [M+H]⁺; HPLC (Method 8)>95% (AUC), t_(R)=6.06 min.

Example 111 Preparation of(E)-1-(3-(3-amino-1-hydroxypropyl)styryl)cyclohexanol

(E)-1-(3-(3-Amino-1-hydroxypropyl)styryl)cyclohexanol was preparedaccording to Scheme 41.

Step 1:

To a −78° C. solution of acetonitrile (1.05 mL, 20 mmol) in anhydrousTHF (25 mL) under argon, was added lithium diisopropylamide (11 mL of a2 M solution in THF, 22 mmol) dropwise. The resulting mixture wasstirred at −78° C. for 15 min. A solution of 3-bromobenzaldehyde (7)(2.78 g, 15 mmol) in anhydrous THF (10 mL) was added dropwise. Thereaction mixture was allowed to warm to room temperature, thenconcentrated under reduced pressure and diluted with EtOAc (75 mL). Thesolution was washed with water (50 mL) and brine (50 mL), dried overNa₂SO₄ and concentrated under reduced pressure. Purification by flashchromatography (20 to 100% EtOAc-hexanes gradient) gave3-(3-bromophenyl)-3-hydroxypropanenitrile (120) as a light yellow oil.Yield (2.75 g, 81%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.60 (t, J=1.6 Hz, 1H),7.46 (ddd, J=7.6, 2.0, 1.2 Hz, 1H), 7.40 (dd, J=7.6, 2.0 Hz, 1H), 7.31(t, J=7.6 Hz, 1H), 6.05 (d, J=4.8 Hz, 1H), 2.94-2.80 (m, 2H).

Step 2:

A mixture of 1-vinylcyclohexanol (120) (0.25 g, 2 mmol), Palladiumacetate (30 mg), 3-(3-bromophenyl)-3-hydroxypropanenitrile (119) (0.45g, 2 mmol) and tetrabutylamonium acetae (1.0 g) was stirred at 90° C.under argon for 18 hours. The reaction mixture was cooled to roomtemperature and partitioned between water and ethyl acetate. The organiclayer was dried over Na₂SO₄ and concentrated under reduced pressure.Purification by flash chromatography (10 to 50% EtOAc-hexanes gradient)gave olefin 121 as a white solid. Yield (0.50 g, 95%): ¹H NMR (400 MHz,DMSO-d₆) δ 7.41 (s, 1H), 7.21-7.30 (m, 3H), 6.20 (d, J=16 Hz, 1H), 6.38(d, J=16 Hz, 1H), 5.90 (d, J=1.6 Hz, 1H), 4.85 (q, J=4.8 Hz, 1H), 4.42(s, 1H), 2.77-2.91 (m, 2H), 1.54-1.68 (m, 2H), 1.38-1.54 (m, 7H),1.19-1.28 (m, 1H).

Step 3:

To a −10° C. solution of olefin 121 (0.35 g, 1.3 mmol) in ether (15 ml)was added LiAlH₄ (X mL of a 2 M in THF, 4.2 mmol) The reaction mixturewas stirred at the temperature for 1.5 hours and the reaction wasquenched by the addition of ice, followed br sat. aqueous Na₂SO₄.Ammonia (3 ml of a 7N solution in MeOH) was added. The mixture was thendiluted with DCM (30 ml). The organic layer was dried over Na₂SO₄ andconcentrated under reduced pressure. Purification by flashchromatography (10 to 15 to 25% 7N NH₃ in MeOH-DCM gradient) gaveExample 111 as colorless oil, Yield (0.20 g, 56%): ¹H NMR (400 MHz,DMSO-d₆) δ 7.33 (s, 1H), 7.20-7.22 (m, 2H), 7.11-7.14 (m, 1H), 6.50 (d,J=16 Hz, 1H), 6.36 (d, J=16 Hz, 1H), 4.63 (t, J=5.4 Hz, 1H), 4.39 (s,1H), 2.54-2.64 (m, 2H), 1.56-1.78 (m, 4H), 1.38-1.56 (m, 7H), 1.19-1.28(m, 1H).

Example 112 ((E)-1-(3-(2-aminoethoxy)styryl)cyclohexanol

(E)-1-(3-(2-Aminoethoxy)styryl)cyclohexanol was prepared according tothe methods used in Examples 31, 11 and 18.

Step 1:

Alkylation of 3-iodophenol with 2-(2-hydroxyethyl)isoindoline-1,3-dionedione following method used in Example 31 gave2-(2-(3-iodophenoxy)ethyl)isoindoline-1,3-dione. Yield (2.8 g, 71%): ¹HNMR (400 MHz, CDCl₃) δ 7.85-7.87 (m, 2H), 7.71-7.73 (m, 2H), 7.22-7.26(m, 3H), 6.94 (t, J=8.4 Hz, 1H), 6.81-6.84 (m, 1H), 4.19 (t, J=5.6 Hz,2H), 4.09 (t, J=5.2 Hz, 2H).

Step 2:

Coupling of 2-(2-(3-iodophenoxy)ethyl)isoindoline-1,3-dione with olefin120 following the method used in Example 111 gave(E)-2-(2-(3-(2-(1-hydroxycyclohexyl)vinyl)phenoxy)ethyl)isoindoline-1,3-dione.Yield (0.50 g, 95%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.81-7.88 (m, 4H), 7.15(t, J=8.0 Hz, 1H), 6.90-6.92 (m, 2H), 6.69-6.71 (m, 1H), 6.64 (d, J=16Hz, 1H), 6.34 (d, J=16 Hz, 1H), 4.38 (s, 1H), 4.19 (t, J=6.0 Hz, 2H),2.93 (t, J=6.0 Hz, 2H), 1.38-1.66 (m, 9H), 1.19-1.26 (m, 1H).

Step 3:

Deprotection of(E)-2-(2-(3-(2-(1-hydroxycyclohexyl)vinyl)phenoxy)ethyl)isoindoline-1,3-dionefollowing the method used in Example 18 gave Example 112 as a colorlessoil. Yield (0.20 g, 56%): ¹H NMR (400 MHz, MeOD) δ 7.19 (t, J=8.0 Hz,1H), 6.96-6.98 (m, 2H), 6.78-6.80 (m, 1H), 6.55 (d, J=16.4 Hz, 1H), 6.33(d, J=16 Hz, 1H), 4.10 (t, J=5.2 Hz, 2H), 2.99 (t, J=5.2 Hz, 2H),1.20-1.78 (m, 12H).

Example 113(E)-1-(3-((1R,2R)-3-amino-1-hydroxy-2-methylpropyl)styryl)cyclohexanol

(E)-1-(3-((1R,2R)-3-Amino-1-hydroxy-2-methylpropyl)styryl)cyclohexanolwas prepared according to Scheme 42.

Step 1.

To a mixture of 3-bromobenzaldehyde (7) (4.16 g, 22.5 mmol),(R)-4-benzyl-3-propionyloxazolidin-2-one (122) (5.111 g, 21.9 mmol) andanhydrous MgCl₂ (0.21 g, 2.23 mmol) in ethyl acetate (40 mL) was addedEt₃N (6.3 mL, 45.2 mmol) and chlorotrimethylsilane (4.3 mL, 34.0 mmol)under argon. The reaction mixture was stirred for 22 hrs at roomtemperature, then filtered through a layer of a silica gel, washing withEtOAc. The filtrate was concentrated under reduced pressure and theresidue was purified by flash chromatography (2 to 25% EtOAc/hexanegradient) to give oxazolidinone 123 as a colorless oil. Yield (9.79 g,91%); ¹H NMR (400 MHz, DMSO-d₆) δ 7.56 (t, J=1.8 Hz, 1H), 7.49 (ddd,J=1.2, 2.0, 7.8 Hz, 1H), 7.40 (dt, J=1.2, 7.6 Hz, 1H), 7.23-7.35 (m,5H), 4.94 (d, J=9.4 Hz, 1H), 4.67-4.75 (m, 1H), 4.30 (t, J=8.6 Hz, 1H),4.12 (dd, J=2.9, 8.8 Hz, 1H), 4.00-4.08 (m, 1H), 3.02 (dd, J=3.1, 13.5Hz, 1H), 2.91 (dd, J=7.4, 13.5 Hz, 1H), 0.78 (d, J=7.0 Hz, 3H), −0.09(s, 9H).

Step 2.

To an ice-cold solution of LiBH₄ (2M in THF, 65 mL, 130 mmol) was addedMeOH (2.6 mL, 64.2 mmol) and the mixture was stirred at 0° C. for 5mins. A solution of oxazolidinone 123 (9.59 g, 19.6 mmol) in anhydrousTHF (170 mL) was added and reaction mixture was stirred at 0° C. for 1.5hrs and then at room temperature for 1.5 hrs. An aqueous solution ofNH₄Cl (25%, 75 mL) was added slowly to reaction mixture for over 1 hrfollowed by addition of EtOAc and stirring was continued at roomtemperature until the mixture became clear. The layers were separatedand the aqueous layer was extracted with EtOAc. The combined organiclayers were washed with saturated brine, dried with anhydrous MgSO₄,filtered and concentrated under reduced pressure. The residue waspurified by flash chromatography (5 to 30% EtOAc/hexane gradient) togive alcohol 124 as colorless oil. Yield (2.97 g, 48%); ¹H NMR (400 MHz,DMSO-d₆) δ 7.38-7.43 (m, 2H), 7.24-7.27 (m. 2H), 4.58 (d, J=6.85 Hz,1H), 4.38 (t, J=5.3 Hz, 1H), 3.32-3.38 (m, 1H), 3.22-3.29 (m, 1H),1.73-1.80 (m, 1H), 0.61 (d, J=6.85 Hz, 3H), −0.05 (s, 9H).

Step 3.

DEAD (1.9 mL, 11.4 mmol) was added to a solution of alcohol 124 (2.97 g,9.36 mmol), phthalimide (1.52 g, 10.3 mmol) and Ph₃P (3.02 g, 11.5 mmol)in anhydrous THF (40 mL) and the mixture was stirred at room temperaturefor 1 hr. The solvent was concentrated under reduced pressure to give anorange residue which was vigorously stirred with 10% EtOAc in hexanes.The triphenylphosphine oxide precipitated and was removed by filtration,washing with 5% EtOAc in hexanes. The filtrate was concentrated underreduced pressure and the residue was purified by flash chromatography (5to 30% EtOAc/hexane gradient) to give bromide 125 as colorless oil.Yield (3.97 g, 95%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.76-7.80 (m, 4H), 7.47(t, J=1.8 Hz, 1H), 7.27-7.35 (m, 2H), 7.22 (t, J=7.8 Hz, 1H), 4.66 (d,J=5.7 Hz, 1H), 3.63 (dd, J=5.7, 13.7 Hz, 1H), 3.38 (dd, J=8.8, 13.5 Hz,1H), 2.24-2.32 (m, 1H), 0.68 (d, J=6.8 Hz, 3H), −0.03 (s, 9H).

Step 4:

Bromide 125 was coupled with alkene 120 following the method used inExample 111. Purification by flash column chromatography (silica gel,30% to 70% EtOAc/hexanes gradient) afforded phthalimide 126 as acolorless oil. Yield (0.281 g, 65%): ¹H NMR (400 MHz, DMSO-d₆) δ7.75-7.81 (m, 4H), 7.33-7.36 (m, 1H), 7.10-7.22 (m, 2H), 6.49 (d, J=16.0Hz, 1H), 6.34 (d, J=16.0 Hz, 1H), 5.33 (d, J=4.3 Hz, 1H), 4.42 (dd,J=4.3, 5.9 Hz, 1H), 4.40 (s, 1H), 3.72 (dd, J=5.5, 13.7 Hz, 1H), 3.42(dd, J=9.4, 13.7 Hz, 1H), 2.20-2.30 (m, 1H), 1.55-1.66 (m, 2H),1.38-1.55 (m, 7H), 1.16-1.26 (m, 1H), 0.65 (d, J=6.9 Hz, 3H).

Step 5:

Deprotection of phthalimide 126 was performed following the method usedin Example 32 except the reaction was heated at 50° C. overnight.Purification by flash column chromatography (silica gel, 50% to 100% of20% 7N NH₃/MeOH in EtOAc/hexanes gradient) gave Example 113 as a whitewaxy solid. Yield (0.154 g, 94%): ¹H NMR (400 MHz, MeOH-d₄) δ 7.35-7.38(m, 1H), 7.23-7.30 (m, 2H), 7.16 (dt, J=1.4, 7.0 Hz, 1H), 6.60 (d,J=16.2 Hz, 1H), 6.35 (d, J=16.2 Hz, 1H), 4.40 (d, J=8.0 Hz, 1H), 2.83(dd, J=5.5, 12.7 Hz, 1H), 2.67 (dd, J=5.9, 12.5 Hz, 1H), 1.79-1.89 (m,1H), 1.47-1.76 (m, 9H), 1.26-1.40 (m, 1H), 0.72 (d, J=6.9 Hz, 3H). ¹³CNMR (100 MHz, MeOH-d₄) δ 144.5, 137.8, 137.6, 128.2, 126.8, 125.7,125.2, 124.6, 78.6, 71.2, 45.3, 42.2, 37.5, 25.5, 21.9, 20.9, 13.9. ESIMS m/z 290.3 [M+H]⁺; HPLC (Method 8) 93% (AUC), t_(R)=3.09 min.

Example 114(E)-1-(3-(3-amino-1-hydroxypropyl)-5-fluorostyryl)cyclohexanol

(E)-1-(3-(3-Amino-1-hydroxypropyl)-5-fluorostyryl)cyclohexanol wasprepared according to the Methods used in Examples 50, 79 and 111.

Step 1:

Alkylation of 3-bromo-5-fluorobenzaldehyde with acetonitrile followingthe method used in Example 50 except KOBu-t was used instead of LDA gave3-(3-bromo-5-fluorophenyl)-3-hydroxypropanenitrile as a light color oil.Yield (2.5 g, 86%): ¹H NMR (400 MHz, CDCl₃) δ 7.34 (bs, 1H), 7.21-7.24(m, 1H), 7.08-7.11 (m, 1H), 5.02 (t, J=6.4 Hz, 1H), 2.75 (d, J=6.4 Hz,2H).

Step 2:

To a solution of 3-(3-bromo-5-fluorophenyl)-3-hydroxypropanenitrile(2.50 g, 8.65 mmol) in THF (16.0 mL) was added borane methyl sulfide(1.20 g, 15.7 mmol) The mixture was heated to reflux for 2 h. Aftercooling to room temperature, saturated aqueous NaHCO₃ (10 mL) was addedand the mixture was stirred for 1 h. The THF was removed under reducedpressure and the remaining aqueous portion was extracted with ethylacetate (2×60 ml). The extract was dried (Na₂SO₄) and concentrated togive 3-amino-1-(3-bromo-5-fluorophenyl)propan-1-ol that was used infollowing reaction without further purification.

Step 3:

Protection of amine following the method used in Example 79 gaveN-(3-(3-bromo-5-fluorophenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamideas a light yellow oil. Yield (1.0 g, 30% in 2 steps): ¹H NMR (400 MHz,DMSO-d₆) δ 9.30 (m, 1H), 7.34-7.38 (m, 2H), 7.15-7.18 (m, 1H), 5.57 (d,J=4.4 Hz, 1H), 4.58-4.61 (m, 1H), 3.16-3.30 (m, 2H), 1.70-1.88 (m, 2H).

Step 4:

Coupling of olefin 120 withN-(3-(3-bromo-5-fluorophenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamidefollowing the method used in Example 111 except the reaction was done in1 hour at 90° C., gave(E)-2,2,2-trifluoro-N-(3-(5-fluoro-3-(2-(1-hydroxycyclohexyl)vinyl)phenyl)-3-hydroxypropyl)acetamideas a light yellow oil. Yield (0.25 g, 54%): ¹H NMR (400 MHz, DMSO-d₆) δ9.32 (m, 1H), 6.94-7.18 (m, 3H), 6.51 (d, J=16 Hz, 1H), 6.43 (d, J=16Hz, 1H), 5.42 (d, J=4.4 Hz, 1H), 4.54-4.59 (m, 1H), 3.18-3.28 (m, 2H),1.73-1.83 (m, 2H), 1.18-1.68 (m, 10H).

Step 4:

Deprotection of(E)-2,2,2-trifluoro-N-(3-(5-fluoro-3-(2-(1-hydroxycyclohexyl)vinyl)phenyl)-3-hydroxypropyl)acetamidefollowing the method used in Example 111 gave Example 115 as a lightyellow oil. Yield (0.065 g, 43%): ¹H NMR (400 MHz, MeOD) δ 7.19 (s 1H),6.99-7.02 (m, 1H), 6.93-6.96 (m, 1H), 6.58 (d, J=16 Hz, 1H), 6.40 (d,J=16 Hz, 1H), 4.72 (t, J=6.0 Hz, 1H), 2.75 (m, 2H), 1.28-1.86 (m, 12H).

Example 115(E)-1-(3-(3-amino-1-hydroxypropyl)-2-fluorostyryl)cyclohexanol

(E)-1-(3-(3-Amino-1-hydroxypropyl)-2-fluorostyryl)cyclohexanol wasprepared according to the methods used in Examples 50, 79 and 111.

Step 1:

Alkylation of 3-bromo-2-fluorobenzaldehyde following the method used inExample 50 gave 3-(3-bromo-2-fluorophenyl)-3-hydroxypropanenitrile as alight colored oil. Yield (1.1 g, 45%): ¹H NMR (400 MHz, CDCl₃) δ 7.54(t, J=7.2 Hz, 2H), 7.11 (td, J=7.6, 0.4 Hz, 1H), 5.37 (dd, J=6.8, 4.4Hz, 1H), 2.73-2.91 (m, 2H).

Step 2;

Reduction of 3-(3-bromo-2-fluorophenyl)-3-hydroxypropanenitrile followedby protection of the amine following the method used in Example 79 gaveN-(3-(3-bromo-2-fluorophenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamideas a light colored oil′ Yield (0.30 g, 45%): ¹H NMR (400 MHz, DMSO-d₆) δ7.54-7.58 (m, 1H), 7.45-7.49 (m, 1H), 7.12-7.17 (m, 1H), 4.86 (dd,J=6.8, 4.4 Hz, 1H), 3.27 (t, J=7.2 Hz, 2H), 1.73-1.86 (m, 2H).

Step 3:

Coupling ofN-(3-(3-bromo-2-fluorophenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamidewith olefin 120 following the method used in Example 111 gave(E)-2,2,2-trifluoro-N-(3-(2-fluoro-3-(2-(1-hydroxycyclohexyl)vinyl)phenyl)-3-hydroxypropyl)acetamideas a light yellow oil. Yield (0.13 g, 55%): ¹H NMR (400 MHz, MeOD) δ7.42 (td, J=7.6, 1.2 Hz, 1H), 7.36 (td, J=7.2, 1.2 Hz, 1H), 7.11 (t,J=8.0 Hz, 1H), 6.76 (d, J=16 Hz, 1H), 6.42 (d, J=16 Hz, 1H), 5.02 (dd,J=6.8, 4.4 Hz, 1H), 3.41 (t, J=7.2 Hz, 2H), 1.73-2.00 (m, 2H), 1.50-1.78(m, 9H), 1.30-1.40 (m, 1H).

Step 4:

Deprotection of(E)-2,2,2-trifluoro-N-(3-(2-fluoro-3-(2-(1-hydroxycyclohexyl)vinyl)phenyl)-3-hydroxypropyl)acetamidefollowing the method used in Example 111 gave Example 114 as a colorlessoil. Yield (0.05 g, 56%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.39 (t, J=6.8 Hz,1H), 7.29 (t, J=6.4 Hz, 1H), 7.09 (t, J=7.6 Hz, 1H), 6.62 (d, J=16 Hz,1H), 6.42 (d, J=16 Hz, 1H), 4.88 (t, J=6.0 Hz, 1H), 2.57 (t, J=7.2 Hz,2H), 1.10-1.68 (m, 12H).

Example 116 (E)-4-(3-(3-amino-1-hydroxypropyl)styryl)heptan-4-ol

(E)-4-(3-(3-Amino-1-hydroxypropyl)styryl)heptan-4-ol was preparedaccording to the Methods used in Example 50, 85 and 111.

Step 1:

Alkylation of 3-bromobenzaldehyde with acetonitrile following the methoddescribed in Example 50 gave 3-(3-bromophenyl)-3-hydroxypropanenitrileas a light yellow oil. Yield (2.75 g, 81%): ¹H NMR (400 MHz, DMSO-d₆) δ7.60 (t, J=1.6 Hz, 1H), 7.46 (ddd, J=7.6, 2.0, 1.2 Hz, 1H), 7.40 (dd,J=7.6, 2.0 Hz, 1H), 7.31 (t, J=7.6 Hz, 1H), 6.05 (d, J=4.8 Hz, 1H),2.94-2.80 (m, 2H).

Step 2:

Reduction of 3-(3-bromophenyl)-3-hydroxypropanenitrile following themethod used in Example 50 gave 3-amino-1-(3-bromophenyl)propan-1-ol as alight green oil. Yield (2.30 g, 84%.) This material was used in the nextstep without further purification. ¹H NMR (400 MHz, DMSO-d₆) δ 7.49 (m,1H), 7.37 (dt, J=7.2, 1.6 Hz, 1H), 7.23-7.31 (m, 2H), 4.66 (t, J=6.8 Hz,1H), 2.61 (m, 2H), 1.61 (q, J=6.8 Hz, 2H).

Step 3:

Protection of 3-amino-1-(3-bromophenyl)propan-1-ol following the methodused in Example 85 except that the reaction mixture was stirred for 3hours, gaveN-(3-(3-bromophenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamide as an oilcontaining ˜15% of2,2,2-trifluoro-N-(3-hydroxy-3-phenylpropyl)acetamide. Yield (1.96 g,60%): ¹H NMR (400 MHz, DMSO-d₆) δ 9.33 (s, 1H), 7.51 (t, J=2.0 Hz, 1H),7.41 (dt, J=7.6, 2.0 Hz, 1H), 7.25-7.32 (m, 2H), 5.46 (d, J=6.4 Hz, 1H),4.55-4.60 (m, 1H), 3.20-3.23 (m, 2H), 1.75-1.82 (m, 2H).

Step 4:

Coupling of 4-vinylheptan-4-ol withN-(3-(3-bromophenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamide followingthe method in Example 111 except the reaction was done in 1 hour at 90°C. gave(E)-2,2,2-trifluoro-N-(3-hydroxy-3-(3-(3-hydroxy-3-propylhex-1-enyl)phenyl)propyl)acetamideas a light color oil. Yield (42 g, 45%): ¹H NMR (400 MHz, MeOD) δ 7.39(s, 1H), 7.26-7.27 (m, 2H), 7.18-7.20 (m, 1H), 6.54 (d, J=16 Hz, 1H),6.23 (d, J=16 Hz, 1H), 4.67 (t, J=6.4 Hz, 1H), 3.37 (t, J=7.2 Hz, 2H),1.92-1.97 (m, 2H), 1.55-1.59 (m, 4H), 1.30-1.46 (m, 4H), 0.91 (t, J=7.6Hz, 6H).

Step 5:

Deprotection of(E)-2,2,2-trifluoro-N-(3-hydroxy-3-(3-(3-hydroxy-3-propylhex-1-enyl)phenyl)propyl)acetamidefollowing the method used in Example 111 gave Example 116 as a colorlessoil. Yield (0.2 g, 63%): ¹H NMR (400 MHz, MeOD) δ 7.39 (s, 1H),7.24-7.26 (m, 2H), 7.18-7.20 (m, 1H), 6.53 (d, J=16 Hz, 1H), 6.23 (d,J=16 Hz, 1H), 4.71 (dd, J=8.0, 5.6 Hz, 1H), 2.68-2.79 (m, 2H), 1.78-1.94(m, 2H), 1.54-1.59 (m, 4H), 1.30-1.46 (m, 4H), 0.91 (t, J=7.6 Hz, 6H).

Example 117 Preparation of(1S,2S)-3-amino-1-(3-((E)-2-(1-hydroxycyclohexyl)vinyl)phenyl)propane-1,2-diol

(1S,2S)-3-amino-1-(3-((E)-2-(1-hydroxycyclohexyl)vinyl)phenyl)propane-1,2-diolwas prepared following the method described in Scheme 43.

Step 1:

To an ice-cold solution of 3-bromobenzaldehyde (7) (3.9 mL, 33.30 mmol)in anhydrous dichloromethane (100 mL) was added(carbethoxymethylene)triphenylphosphorane (11.65 g, 33.44 mmol) and thereaction mixture was stirred at 0° C. for 5 min, then allowed to warm toroom temperature over 30 min. Then the reaction mixture was concentratedunder reduced pressure. White sticky solid was resuspended in 5%EtOAc/hexanes, stirred for 10 min at room temperature and then filtered.Filter cake was washed with hexanes, and the filtrate was concentratedunder reduced pressure. The residue was purified by flash columnchromatography (silica gel, hexanes to 10% EtOAc/hexanes gradient) togive allyl ester 127 as white solid. Yield (7.63 g, 90%). ¹H NMR (400MHz, DMSO-d₆) δ 7.95 (t, J=1.8 Hz, 1H), 7.70-7.72 (m, 1H), 7.59 (d,J=16.4 Hz, 1H), 7.58 (ddd, J=1.0, 2.0, 8.0 Hz, 1H), 7.35 (t, J=7.8 Hz,1H), 6.69 (d, J=16.0 Hz, 1H), 4.17 (q, J=7.2 Hz, 2H), 1.23 (t, J=7.0 Hz,3H).

Step 2.

To an ice-cold solution of ester 127 (7.63 g, 29.9 mmol) in diethylether (100 mL) was added a solution of diisobutyl aluminum hydride(DIBAL-H, 60 mL of a 1.0 M solution in CH₂Cl₂, 60.0 mmol). The reactionwas stirred at 0° C. for 30 min after which aqueous solution of NaHSO₄(2M, 42 mL) was added and the mixture was stirred for 1.5 hrs whilewarming to room temperature. Anhydrous MgSO₄ was added to the stirredreaction mixture, and after 30 min mixture was filtered, filtrate cakewashed excessively with EtOAc, and filtrate was concentrated underreduced pressure to afford alcohol 128 as a colorless oil. Yield (6.42g, quant.). ¹H NMR (400 MHz, DMSO-d₆) δ 7.60 (t, J=1.8 Hz, 1H), 7.40(dt, J=1.2, 7.6 Hz, 1H), 7.38 (ddd, J=1.0, 2.0, 8.0 Hz, 1H), 7.25 (t,J=7.6 Hz, 1H), 6.48-6.54 (m, 1H), 6.43 (dt, J=4.3, 16.0 Hz, 1H), 4.88(t, J=5.5 Hz, 1H), 4.08-4.12 (m, 2H).

Step 3.

Acetylation of alcohol 128 following the method used in Example 51 afterflash column chromatography (silica gel, 5% to 10% EtOAc/hexanesgradient) gave acetate 129 as a colorless oil. Yield (2.71 g, 89%). ¹HNMR (400 MHz, DMSO-d₆) δ 7.66 (t, J=1.8 Hz, 1H), 7.40-7.46 (m, 2H), 7.27(t, J=7.8 Hz, 1H), 6.58-6.65 (m, 1H), 6.42 (dt, J=5.9, 16.0 Hz, 1H),4.66 (dd, J=1.4, 5.9 Hz, 1H), 2.04 (s, 3H).

Step 4.

A mixture of allyl acetate 129 (2.71 g, 10.6 mmol), sodium azide (0.787g, 12.1 mmol), water (20 mL) and THF (50 mL) was degassed by bubblingargon for 3 min and tris-dibenzylideneacetonyl-bis-palladium-chloroformadduct (0.158 g, 0.17 mmol), diphenylphosphinoferrocene (0.1773 g, 0.32mmol) were added to the reaction mixture. Air was evacuated by applyingvacuum/argon 3× and then the reaction mixture was heated at 60° C. underargon for 4 hrs. The reaction mixture was concentrated under reducedpressure, water added to the residue and the product was extracted twicewith hexanes, hexane layers were washed with saturated brine, dried withanhydrous MgSO₄, filtered and the filtrate was concentrated underreduced pressure. The residue was purified by flash columnchromatography (silica gel, 5% to 30% EtOAc/hexanes gradient) to giveallyl azide 130 as a colorless oil. Yield (1.90 g, 75%). ¹H NMR (400MHz, DMSO-d₆) δ 7.69 (t, J=1.8 Hz, 1H), 7.42-7.48 (m, 2H), 7.28 (t,J=7.8 Hz, 1H), 6.62-6.68 (m, 1H), 6.45 (dt, J=6.3, 15.8 Hz, 1H), 4.02(dd, J=1.2, 6.3 Hz, 1H).

Step 5.

To a 100-ml round bottomed flask was placed H₂O (19 mL) and tert-BuOH(19 mL) followed by AD-mix-β (5.61 g). The mixture was stirred at roomtemperature for 10 min after which MeSO₂NH₂ (0.36 g, 3.79 mmol) wasadded. The reaction mixture was cooled to 0° C., allyl azide 130 (0.90g, 3.78 mmol) was added and stirred at 0° C. for 24 hrs. Na₂SO₃ (6.30 g)was added and the mixture was stirred for another hour after which EtOAc(50 mL) was added followed by sat. NaCl (50 mL). Layers were separatedand aqueous layer extracted with EtOAc (3×25 mL). Combined organiclayers were washed with brine (50 mL), dried with anhydrous MgSO₄ andfiltered. Filtrate was concentrated under reduced pressure and theresidue was purified by flash column chromatography (silica gel, 10% to90% EtOAc/hexanes gradient) to give azido diol 131 as a thick colorlessoil. Yield (1.02 g, 99%). ¹H NMR (400 MHz, DMSO-d₆) δ 7.50 (t, J=1.6 Hz,1H), 7.40 (ddd, J=1.2, 2.0, 7.6 Hz, 1H), 7.29-7.33 (m, 1H), 7.25 (t,J=7.6 Hz, 1H), 5.52 (d, J=5.1 Hz, 1H), 5.26 (d, J=5.9 Hz, 1H), 4.51 (t,J=4.7 Hz, 1H), 3.15 (dd, J=3.3, 12.5 Hz, 1H), 3.02 (dd, J=8.0, 12.7 Hz,1H).

Step 6.

A mixture of azido diol 131 (0.826 g, 3.037 mmol), triphenylphosphine(0.84 g, 3.20 mmol), THF (10 mL), water (0.2 mL) and ethyltrifluoroacetate (1 mL) was heated at 50° C. for 5 hrs, thenconcentrated under reduced pressure. The residue was purified by flashcolumn chromatography (silica gel, 20% to 90% EtOAc/hexanes gradient) togive trifluoroacetamide 132 as white solid. Yield 0.73 g, 70%). ¹H NMR(400 MHz, DMSO-d₆) δ 9.21 (t, J=5.3 Hz, 1H), 7.52 (t, J=1.6 Hz, 1H),7.40 (ddd, J=1.2, 2.0, 7.8 Hz, 1H), 7.30-7.33 (m, 1H), 7.25 (t, J=7.6Hz, 1H), 5.48 (d, J=5.1 Hz, 1H), 5.00 (d, J=5.9 Hz, 1H), 4.51 (t, J=4.7Hz, 1H), 3.70-3.76 (m, 1H), 3.24 (dt, J=4.9, 13.3 Hz, 1H), 2.98 (ddd,J=5.7, 8.8, 13.3 Hz, 1H).

Step 7.

Coupling of bromide 132 with olefin 120 was performed following themethod used in Example 111 except that reaction was heated at 90° C. for5 hrs. Purification by flash column chromatography (silica gel, 20% to70% EtOAc/hexanes gradient) afforded alkene 133 as white foam. Yield(0.2128 g, 78%). ¹H NMR (400 MHz, DMSO-d₆) δ 9.19 (t, J=5.3 Hz, 1H),7.35-7.37 (m, 1H), 7.20-7.26 (m, 2H), 7.13-7.18 (m, 1H), 6.51 (d, J=16.2Hz, 1H), 6.33 (d, J=16.0 Hz, 1H), 5.33 (d, J=4.9 Hz, 1H), 4.94 (d, J=5.7Hz, 1H), 4.46 (t, J=4.8 Hz, 1H), 4.41 (s, 1H), 3.70-3.76 (m, 1H), 3.18(dt, J=4.5, 13.3 Hz, 1H), 2.99 (ddd, J=6.1, 8.8, 14.3 Hz, 1H), 1.54-1.66(m, 2H), 1.36-1.54 (m, 7H), 1.18-1.26 (m, 1H).

Step 8.

N-((2S,3S)-2,3-dihydroxy-3-(3-((E)-2-(1-hydroxycyclohexyl)vinyl)phenyl)propyl)-2,2,2-trifluoroacetamide(133) was deprotected according to the method used in Example 79 exceptthat three equivalents of K₂CO₃ were used in a MeOH:H₂O (2:1) mixtureand the reaction mixture was heated at 50° C. for 5 hrs. Following thereaction, reaction mixture was concentrated under reduced pressure,resuspended in EtOAc/EtOH and purified by flash chromatography using agradient of 50% 7N NH₃/MeOH in EtOAc/hexanes to give Example 117 as acolorless oil. Yield (0.118 g, 74%). ¹H NMR (400 MHz, MeOH-d₄) δ7.42-7.44 (m, 1H), 7.30 (dt, J=1.6, 7.6 Hz, 1H), 7.27 (t, J=7.4 Hz, 1H),7.21 (dt, J=1.6, 7.2 Hz, 1H), 6.60 (d, J=16.0 Hz, 1H), 6.36 (d, J=16.0Hz, 1H), 4.50 (d, J=5.9 Hz, 1H), 3.62-2.70 (m, 1H), 2.51-2.58 (m, 2H),1.66-1.77 (m, 2H), 1.48-1.66 (m, 7H), 1.28-1.40 (m, 1H). ¹³C NMR (100MHz, MeOH-d₄) δ 142.3, 137.9, 137.7, 128.3, 126.7, 125.7, 125.6, 124.6,76.0, 75.8, 71.2, 43.6, 37.5, 25.5, 21.9, 20.9. ESI MS m/z 292.3 [M+H]⁺;HPLC (Method 9) 97% (AUC), t_(R)=4.73 min.

Example 118 Preparation of(1R,2R)-3-amino-1-(3-((E)-2-(1-hydroxycyclohexyl)vinyl)phenyl)propane-1,2-diol

N-((2S,3S)-2,3-dihydroxy-3-(3-((E)-2-(1-hydroxycyclohexyl)vinyl)phenyl)propyl)-2,2,2-trifluoroacetamidewas prepared according to the method used in Example 117.

Step 5.

Dihydroxylation of ally azide 130 was conducted using AD-mix-α to give(1R,2R)-3-azido-1-(3-bromophenyl)propane-1,2-diol. Yield (0.966 g, 96%).¹H NMR (400 MHz, DMSO-d₆) δ 7.50 (t, J=1.6 Hz, 1H), 7.40 (ddd, J=1.2,2.0, 7.6 Hz, 1H), 7.29-7.33 (m, 1H), 7.25 (t, J=7.6 Hz, 1H), 5.52 (d,J=5.1 Hz, 1H), 5.26 (d, J=5.9 Hz, 1H), 4.51 (t, J=4.7 Hz, 1H), 3.15 (dd,J=3.3, 12.5 Hz, 1H), 3.02 (dd, J=8.0, 12.7 Hz, 1H).

Step 6.

Reduction and protection of(1R,2R)-3-azido-1-(3-bromophenyl)propane-1,2-diol gaveN-((2R,3R)-3-(3-bromophenyl)-2,3-dihydroxypropyl)-2,2,2-trifluoroacetamideas a white solid. Yield 0.66 g, 69%). ¹H NMR (400 MHz, DMSO-d₆) δ 9.21(t, J=5.3 Hz, 1H), 7.52 (t, J=1.6 Hz, 1H), 7.40 (ddd, J=1.2, 2.0, 7.8Hz, 1H), 7.30-7.33 (m, 1H), 7.25 (t, J=7.6 Hz, 1H), 5.48 (d, J=5.1 Hz,1H), 5.00 (d, J=5.9 Hz, 1H), 4.51 (t, J=4.7 Hz, 1H), 3.70-3.76 (m, 1H),3.24 (dt, J=4.9, 13.3 Hz, 1H), 2.98 (ddd, J=5.7, 8.8, 13.3 Hz, 1H).

Step 7.

Coupling ofN-((2R,3R)-3-(3-bromophenyl)-2,3-dihydroxypropyl)-2,2,2-trifluoroacetamidewith olefin 120 gaveN-((2R,3R)-2,3-dihydroxy-3-(3-((E)-2-(1-hydroxycyclohexyl)vinyl)phenyl)propyl)-2,2,2-trifluoroacetamideas a brownish foam. Yield (0.1958 g, 82%). ¹H NMR (400 MHz, DMSO-d₆) δ9.19 (t, J=5.3 Hz, 1H), 7.35-7.37 (m, 1H), 7.20-7.26 (m, 2H), 7.13-7.18(m, 1H), 6.51 (d, J=16.2 Hz, 1H), 6.33 (d, J=16.0 Hz, 1H), 5.33 (d,J=4.9 Hz, 1H), 4.94 (d, J=5.7 Hz, 1H), 4.46 (t, J=4.8 Hz, 1H), 4.41 (s,1H), 3.70-3.76 (m, 1H), 3.18 (dt, J=4.5, 13.3 Hz, 1H), 2.99 (ddd, J=6.1,8.8, 14.3 Hz, 1H), 1.54-1.66 (m, 2H), 1.36-1.54 (m, 7H), 1.18-1.26 (m,1H).

Step 8.

Deprotection ofN-((2S,3S)-2,3-dihydroxy-3-(3-((E)-2-(1-hydroxycyclohexyl)vinyl)phenyl)propyl)-2,2,2-trifluoroacetamidegave Example 118 as a colorless oil. Yield (0.16 g, quant.). ¹H NMR (400MHz, MeOH-d₄) δ 7.42-7.44 (m, 1H), 7.30 (dt, J=1.6, 7.6 Hz, 1H), 7.27(t, J=7.4 Hz, 1H), 7.21 (dt, J=1.6, 7.2 Hz, 1H), 6.60 (d, J=16.0 Hz,1H), 6.36 (d, J=16.0 Hz, 1H), 4.50 (d, J=5.9 Hz, 1H), 3.62-2.70 (m, 1H),2.51-2.58 (m, 2H), 1.66-1.77 (m, 2H), 1.48-1.66 (m, 7H), 1.28-1.40 (m,1H). ¹³C NMR (100 MHz, MeOH-d₄) δ 142.3, 137.9, 137.7, 128.3, 126.7,125.7, 125.6, 124.6, 76.0, 75.8, 71.2, 43.6, 37.5, 25.5, 21.9, 20.9. ESIMS m/z 292.3 [M+H]⁺; HPLC (Method 9) 96% (AUC), t_(R)=4.73 min.

Example 119 Preparation of(S,E)-1-(3-(1-aminopropan-2-yloxy)styryl)cyclohexanol

(S,E)-1-(3-(1-aminopropan-2-yloxy)styryl)cyclohexanol was preparedfollowing the method shown in scheme 44:

Step 1:

Diethylazodicarboxylate (17.4 g, 100 mmol) was added slowly to asolution of phenol 134 (18.5 g, 84 mmol), alcohol 135 (14.73 g, 84mmol), and triphenyl phosphine (26.2 g, 100 mmol) in THF (200 mL) at 0°C. under argon. The reaction was allowed to warm to room temperature andstirred for 2 hours, then heated to 80° C. for 6 hours. The reaction wasconcentrated under reduced pressure, then triturated with diethyl etherand the resulting white solids removed by filtration. The filtrate wasconcentrated under reduce pressure and the residue was partitionedbetween ethyl acetate and 1N NaOH. The organic layers were combined,washed with brine, and concentrated under reduced pressure. Purificationby flash chromatography (silica gel, eluent 5-15% ethylacetate/hexanesgradient) gave carbamate 136 as an impure yellow oil, which was carriedon to the next step without further purification. Yield (17.3 g, 54%).

Step 2:

HCl (12 mL of a 4.8 M solution in iPrOH, 56 mmol) was added to asolution of carbamate 136 (10 g, 28 mmol) in ethyl acetate (25 mL).After stirring for 1 h, the product was collected by filtration anddried under reduced pressure, to give 137 hydrochloride as a white solidwhich was used in the next step without purification. Yield (2.9 g,30%): ¹H NMR (400 MHz, DMSO-d₆) δ 8.27 (brs, 3H), 7.24-7.28 (m, 2H),6.98-7.12 (m, 2H), 4.68 (m, 1H), 2.90-3.10 (m, 2H), 1.22 (d, 3H).

Step 3:

Protection of 137 hydrochloride with ethyltrifluoroacetate following themethod used in Example 79 gave trifluoroamide 138 as a yellow oil. Yield(3.4 g, quantitative): ¹H NMR (400 MHz, CDCl₃) δ 7.29-7.33 (m, 1H),7.24-7.26 (m, 1H), 6.99 (t, J=8.0 Hz, 1H), 6.83-6.87 (m, 1H), 6.75 (brs,1H), 4.45-4.55 (m, 1H), 3.52-3.53 (m, 1H), 3.40-3.50 (m, 1H), 1.29 (dJ=6.4 Hz, 3H).

Step 4:

Heck coupling of trifluoroamide 138 with 1-vinylcyclohexanol (120)following the method used in Example 111, gave alkene 139 as a yellowglassy oil. Yield (0.286 g, 80%): ¹H NMR (400 MHz, CDCl₃) δ 7.22 (t,J=8.0 Hz, 1H), 6.98-7.04 (m, 1H), 6.90-6.93 (m, 1H), 6.73-6.78 (m, 2H),6.57 (d, J=16 Hz, 1H), 6.32 (d, J=16 Hz, 1H), 4.50-4.59 (m, 1H),3.72-3.80 (m, 1H), 3.40-3.49 (m, 1H), 1.50-1.74 (m, 10H), 1.32-1.38 (m,1H), 1.30 (d, J=6.4 Hz, 3H).

Step 5:

Deprotection of styrene 139 following the method used in Example 85 gaveExample 119 as a colorless oil. Yield (0.154 g, 72%): ¹H NMR (400 MHz,CDCl₃) δ 7.20 (t, J=8.0 Hz, 1H), 6.93-6.98 (m, 2H), 6.76-6.80 (m, 1H),6.57 (d, J=16 Hz, 1H), 6.31 (d, J=16 Hz, 1H), 4.32-4.42 (m, 1H), 2.88(d, J=5.6 Hz, 2H), 1.50-1.74 (m, 12H), 1.28-1.37 (m, 1H), 1.26 (d, J=6.0Hz, 3H); ESI MS m/z 276.3 [M+H].

Example 120 Preparation of(E)-1-(5-(3-amino-1-hydroxypropyl)-2-methoxystyryl)cyclohexanol

(E)-1-(5-(3-Amino-1-hydroxypropyl)-2-methoxystyryl)cyclohexanol wasprepared following the methods used in Example 114:

Step 1:

Alkylation of 3-bromo-4-methoxybenzaldehyde with acetonitrile followingthe method used in Example 115 gave3-(3-bromo-4-methoxyphenyl)-3-hydroxypropanenitrile as a pale orangeoil. Yield (10.32 g, 96%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.58 (d, J=2.0Hz, 1H), 7.35 (dd, J=8.8, 2.0 Hz, 1H), 7.07 (d, J=8.8 Hz, 1H), 5.93 (d,J=4.4 Hz, 1H), 4.85-4.81 (m, 1H). 3.81 (s, 3H), 2.86 (ABd, J=16.4, 4.8Hz, 1H), 2.79 (ABd, J=16.8, 6.8 Hz, 1H).

Step 2:

Reduction of 3-(3-bromo-4-methoxyphenyl)-3-hydroxypropanenitrile withBH₃—S(CH₃)₂ followed by protection of the amine following the method inExample 114 gaveN-(3-(3-bromo-4-methoxyphenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamideas an orange oil. Yield (5.76 g, 40%): ¹H NMR (400 MHz, DMSO-d₆) δ 9.31(bs, 1H), 7.49 (d, J=2.0 Hz, 1H), 7.26 (dd, J=8.8, 2.0 Hz, 1H), 5.32 (d,J=4.8 Hz, 1H), 4.53-4.49 (m, 1H), 3.80 (s, 3H), 3.24-3.15 (m, 2H),1.79-1.72 (m, 2H).

Step 3:

Coupling of olefin 120 withN-(3-(3-bromo-4-methoxyphenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamidefollowing the method used in Example 114 gave(E)-2,2,2-trifluoro-N-(3-hydroxy-3-(3-(2-(1-hydroxycyclohexyl)vinyl)-4-methoxyphenyl)propyl)acetamideas a light yellow oil. Yield (0.24 g, 28%): ¹H NMR (400 MHz, DMSO-d₆) δ9.32 (t, J=5.2 Hz, 1H), 7.38 (d, J=2.0 Hz, 1H), 7.12 (dd, J=8.4, 2.0 Hz,1H), 6.89 (d, J=8.4 Hz, 1H), 6.80 (d, J=16.4 Hz, 1H), 6.28 (d, J=16.4Hz, 1H), 5.19 (d, J=4.8 Hz, 1H), 4.52-4.48 (m, 2H), 4.38 (s, 1H), 3.75(s, 3H), 3.24-3.19 (m, 2H), 1.80-1.75 (m, 2H), 1.65-1.39 (m, 9H),1.25-1.19 (m, 1H).

Step 4:

Deprotection of(E)-2,2,2-trifluoro-N-(3-hydroxy-3-(3-(2-(1-hydroxycyclohexyl)vinyl)-4-methoxyphenyl)propyl)acetamidefollowing the method used in Example 114 gave Example 120 as an offwhite solid. Yield (0.121 g, 68%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.36 (d,J=4.0 Hz, 1H), 7.10 (dd, J=8.4, 4.0 Hz, 1H), 6.87 (d, J=8.4 Hz, 1H),6.79 (d, J=16.4 Hz, 1H), 6.27 (d, J=16.4 Hz, 1H), 4.59-4.56 (m, 1H),4.37 (bs, 1H), 3.74 (s, 3H), 2.65-2.53 (m, 2H), 1.67-1.38 (m, 11H),1.25-1.17 (m, 1H).

Example 121 Preparation of(E)-1-(3-(3-amino-1-hydroxypropyl)-4-chlorostyryl)cyclohexanol

(E)-1-(3-(3-amino-1-hydroxypropyl)-4-chlorostyryl)cyclohexanol wasprepared following the methods used in Example 114.

Step 1:

Alkylation of 5-bromo-2-chlorobenzaldehyde with acetonitrile followingthe method used in Example 114 gave3-(5-bromo-2-chlorophenyl)-3-hydroxypropanenitrile as a pale yellowliquid. Yield (4.42 g, 75%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.74 (d, J=2.8Hz, 1H), 7.53 (dd, J=8.8, 2.8 Hz, 1H), 7.39 (d, J=8.8 Hz, 1H), 6.30 (d,J=4.8 Hz, 1H), 5.13-5.09 (m, 1H), 2.96 (ABd, J=16.8, 4.8 Hz, 1H), 2.83(ABd, J=17.0, 6.0 Hz, 1H).

Step 2:

Reduction of 3-(5-bromo-2-chlorophenyl)-3-hydroxypropanenitrile withBH₃-THF followed by protection of the amine following the method inExample 114 gaveN-(3-(5-bromo-2-chlorophenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamideas an orange oil. Yield (2.6 g, 43%): ¹H NMR (400 MHz, DMSO-d₆) δ 9.42(bs, 1H), 7.67 (d, J=2.4 Hz, 1H), 7.45 (dd, J=8.8, 2.4 Hz, 1H), 7.33 (d,J=8.8 Hz, 1H), 5.64 (d, J=4.4 Hz, 1H), 3.33-3.29 (m, 2H), 1.96-1.80 (m,1H), 1.68-1.59 (m, 1H).

Step 3:

Coupling of olefin 120 withN-(3-(5-bromo-2-chlorophenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamidefollowing the method used in Example 114 except the reaction was donefor 16 hr at 90° C., gave(E)-N-(3-(2-chloro-5-(2-(1-hydroxycyclohexyl)vinyl)phenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamideas a light yellow oil. Yield (0.30 g, 70%): ¹H NMR (400 MHz, DMSO-d₆) δ9.41 (t, J=5.6 Hz, 1H), 7.28-7.26 (m, 2H). 6.51 (d, J=16.0 Hz, 1H), 6.37(d, J=16.0 Hz, 1H), 5.49 (d, J=4.4 Hz, 1H), 4.88-4.86 (m, 1H), 4.34 (bs,1H), 3.34-3.29 (m, 2H), 1.87-1.80 (m, 1H), 1.70-1.39 (m, 10H), 1.25-1.19(m, 1H).

Step 4:

Deprotection of(E)-N-(3-(2-chloro-5-(2-(1-hydroxycyclohexyl)vinyl)phenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamidefollowing the method used in Example 114 gave Example 121 as an offwhite solid. Yield (0.145 g, 64%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.58 (s,1H), 7.27-7.23 (m, 2H), 6.50 (d, J=16.4 Hz, 1H), 6.35 (d, J=16.4 Hz,1H), 4.96-4.93 (m, 1H), 4.44 (bs, 1H), 2.70-2.63 (m, 2H), 2.47-1.38 (m,11H), 1.25-1.16 (m, 1H).:

Example 122 Preparation of(E)-1-(3-(3-amino-1-hydroxypropyl)-4-methylstyryl)cyclohexanol

(E)-1-(3-(3-Amino-1-hydroxypropyl)-4-methylstyryl)cyclohexanol wasprepared according to the Methods used in Example 115.

Step 1:

Alkylation of 5-bromo-2-methylbenzaldehyde with acetonitrile followingthe method used in Example 114 gave3-(5-bromo-2-methylphenyl)-3-hydroxypropanenitrile as a pale yellow oil.Yield (3.33 g, 86%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.61 (d, J=2.0 Hz, 1H),7.35 (dd, J=8.0, 2.0 Hz, 1H), 7.09 (d, J=8.0 Hz, 1H), 5.96 (d, J=4.4 Hz,1H), 5.04-5.00 (m, 1H), 2.88 (ABd, J=16.8, 4.4 Hz, 1H), 2.77 (ABd,J=16.8, 6.4 Hz, 1H), 2.23 (s, 3H).

Step 2:

Reduction of 3-(3-bromo-2-methylphenyl)-3-hydroxypropanenitrile withBH₃—S(CH₃)₂ followed by protection of the amine following the method inExample 115 gaveN-(3-(3-bromo-2-methylphenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamideas a pale yellow oil. Yield (3.25 g, 69%): ¹H NMR (400 MHz, DMSO-d₆) δ9.38 (bs, 1H), 7.53 (d, J=2.4 Hz, 1H), 7.28 (dd, J=8.0, 2.4 Hz, 1H),7.05 (d, J=8.0 Hz, 1H), 4.73-4.70 (m, 1H), 3.36-3.26 (m, 2H), 2.17 (s,3H), 1.79-1.71 (m, 1H), 1.68-1.59 (m, 1H).

Step 3:

Coupling of olefin 120 withN-(3-(3-bromo-2-methylphenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamidefollowing the method used in Example 115 except the reaction was donefor 16 hr at 90° C., gave(E)-2,2,2-trifluoro-N-(3-hydroxy-3-(5-(2-(1-hydroxycyclohexyl)vinyl)-2-methylphenyl)propyl)acetamideas a clear oil. Yield (0.372 g, 47%): ¹H NMR (400 MHz, DMSO-d₆) δ 9.38(t, J=5.2 Hz, 1H), 7.44 (d, J=2.0 Hz, 1H), 7.12 (dd, J=8.0, 2.0 Hz, 1H),7.02 (d, J=8.0 Hz, 1H), 6.46 (d, J=16.4 Hz, 1H), 6.27 (d, J=16.4 Hz,1H), 5.19 (d, J=3.6 Hz, 1H), 4.74-4.72 (m, 1H), 4.37 (bs, 1H), 3.33-3.28(m, 2H), 2.19 (s, 3H), 1.80-1.38 (m, 11H), 1.25-1.16 (m, 1H).

Step 4:

Deprotection of(E)-2,2,2-trifluoro-N-(3-hydroxy-3-(5-(2-(1-hydroxycyclohexyl)vinyl)-2-methylphenyl)propyl)acetamidefollowing the method used in Example 115 gave Example 122 as a paleyellow solid. Yield (0.145 g, 53%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.44 (d,J=1.6 Hz, 1H), 7.09 (dd, J=8.0, 1.6 Hz, 1H), 6.99 (d, J=8.0 Hz, 1H),6.46 (d, J=16.0 Hz, 1H), 6.25 (d, J=16.0 Hz, 1H), 4.84-4.81 (m, 1H),4.47 (bs, 1H), 2.73-2.61 (m, 2H), 2.21 (s, 3H), 1.73-1.38 (m, 11H),1.25-1.16 (m, 1H).

Example 123 Preparation of(E)-1-(3-(3-amino-1-hydroxypropyl)-5-methylstyryl)cyclohexanol

(E)-1-(3-(3-Amino-1-hydroxypropyl)-5-methylstyryl)cyclohexanol wasprepared according to the Methods used in Examples 50, 79 and 111.

Step 1:

Alkylation of 3-bromo-5-methylbenzaldehyde following the method used inExample 50 gave 3-(3-bromo-5-methylphenyl)-3-hydroxypropanenitrile,which was used directly in the next step.

Step 2:

Reduction of 3-(3-bromo-5-methylphenyl)-3-hydroxypropanenitrile usingborane methyl sulfide following the method used in Example 79 gave3-amino-1-(3-bromo-5-methylphenyl)propan-1-ol which was used directly inthe next step.

Step 3:

Treatment of 3-amino-1-(3-bromo-5-methylphenyl)propan-1-ol with ethyltrifluoroacetate following the method used in Example 79 gaveN-(3-(3-bromo-5-methylphenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamideas a light colored oil. Yield (0.38 g, 22%, 3 steps): ¹H NMR (400 MHz,CDCl₃) δ 7.29 (s, 1H), 7.26 (s, 1H), 7.05-7.07 (m, 1H), 4.80 (dd, J=8.8,4.0 Hz, 1H), 3.62-3.75 (m, 1H), 3.36-3.44 (m, 1H), 2.33 (s, 3H),1.90-2.0 (m, 2H).

Step 4:

Coupling ofN-(3-(3-bromo-5-methylphenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamidewith olefin 120 following the method used in Example 111 gave(E)-2,2,2-trifluoro-N-(3-hydroxy-3-(3-(2-(1-hydroxycyclohexyl)vinyl)-5-methylphenyl)propyl)acetamideas a light yellow oil. Yield (0.28 g, 65%): ¹H NMR (400 MHz, DMSO-d₆) δ9.34 (t, J=4.8 Hz, 1H), 7.12 (s, 1H), 7.05 (s, 1H), 7.96 (s, 1H), 6.46(d, J=16 Hz, 1H), 6.32 (d, J=16 Hz, 1H), 5.24 (d, J=4.8 Hz, 1H), 4.50(q, J=4.8 Hz, 1H), 4.38 (s, 1H), 3.22 (q, J=7.6 Hz, 2H), 2.25 (s, 3H),1.65-1.70 (m, 2H), 1.54-1.68 (m, 2H), 1.37-1.53 (m, 7H), 1.18-1.26 (m,1H).

Step 5:

Deprotection of(E)-2,2,2-trifluoro-N-(3-hydroxy-3-(3-(2-(1-hydroxycyclohexyl)vinyl)-5-methylphenyl)(E)-1-(3-(3-Amino-1-hydroxypropyl)-5-methylstyryl)cyclohexanol. The freebase was dissolved in ethyl acetate (10 ml) and HCl.EtOH (6.95 M, 2 ml)added. The mixture was concentrated under reduced pressure. To theresidue was added 30% ethyl acetate/hexane (10 ml) and mixture wassonicated. Collection of the solid by filtration followed by drying gaveExample 128 hydrochloride as a light color. Yield (0.07 g, 29%): ¹H NMR(400 MHz, DMSO-d₆) δ 7.70 (bs, 3H), 7.18 (s, 1H), 7.14 (s, 1H), 6.96 (s,1H), 6.79 (d, J=16.4 Hz, 1H), 6.38 (d, J=16.4 Hz, 1H), 5.91 (bs, 1H),5.48 (d, J=2.8 Hz, 1H), 4.58-4.64 (m, 1H), 2.76-2.85 (m, 2H), 2.27 (s,3H), 2.08-2.21 (m, 5H), 1.75-1.88 (m, 2H), 1.51-1.68 (m, 5H).

Example 124 Preparation of(1S,2R)-3-amino-1-(3-((E)-2-(1-hydroxycyclohexyl)vinylphenyl)propane-1,2-diol

(1S,2R)-3-amino-1-(3-((E)-2-(1-hydroxycyclohexyl)vinyl)phenyl)propane-1,2-diolwas prepared following the method described in Scheme 45.

Step 1:

To an ice-cold solution of 3-bromobenzaldehyde (7) (3.2 mL, 27.3 mmol)in anhydrous diethyl ether (50 mL) was slowly added a fresh solution ofvinyl magnesium bromide (30.0 mL of a 1.0 M solution in THF, 30 mmol).The reaction mixture was stirred at 0° C. for 20 min, after whichaqueous solution of NH₄Cl (25%, 50 mL) was added. The mixture wasallowed to warm to room temperature, layers were separated and aqueouslayer was extracted with hexane. Combined organic layers were washedwith brine, concentrated under reduced pressure and purified by flashcolumn chromatography (silica gel, 5% to 300% EtOAc/hexanes gradient) togive allyl alcohol 141 as a colorless oil. Yield (4.22 g, 73%). ¹H NMR(400 MHz, DMSO-d₆) δ 7.47-7.49 (m, 1H), 7.40 (dt, J=1.8, 7.4 Hz, 1H),7.24-7.32 (m, 2H), 5.85-5.94 (m, 1H), 5.61 (d, J=4.5 Hz, 1H), 5.24 (dt,J=1.8, 17.0 Hz, 1H), 5.00-5.07 (m, 2H).

Step 2.

To a cold (−23° C.) mixture of powdered 4 Å molecular sieves (6.4 g) andtitanium tetraisopropoxide (5.5 mL, 18.8 mmol) in anhydrous CH₂Cl₂ (110mL) was added L-(+)-diisopropyl tartrate (DIPT, 4.7 mL, 22.49 mmol)under inert atmosphere. The reaction mixture was stirred at −20° C. anda solution of allyl alcohol 141 (4.0 g, 18.8 mmol) in anhydrous CH₂Cl₂(80 mL) was added over 5 mins After the reaction mixture was stirred at−20° C. for 30 min, tert-butyl hydroperoxide solution (5.0-6.0 M innonane, 2 mL, ca 10.0 mmol) was added. The reaction mixture was stirredat −20° C. for 7.5 nrs, kept at −20° C. overnight, stirred at −20° C.for another 7 hrs and left at −20° C. and then kept at −20° C. for 43hrs. An aqueous solution of L-tartaric acid (10%, 110 mL) was added tothe reaction mixture, the mixture was stirred for 10 min at roomtemperature, then saturated aqueous solution of Na₂SO₄ (20 mL) wasadded. The mixture was stirred vigorously for 1 h at room temperature,layers were separated. Aqueous layer was extracted with diethyl ether,then with EtOAc. Combined organic layers were washed with brine, driedover anhydrous NaSO₄, filtered and the filtrate concentrated underreduced pressure. The residue was purified by flash columnchromatography (silica gel, 30% to 70% EtOAc/hexanes gradient) to give amixture of epoxide 142 and DIPT (1:1 molar ratio) as a colorless oil andunreacted (S)-1-(3-bromophenyl)prop-2-en-1-ol 143 (2.16 g) as acolorless oil. Crude epoxide 142 was re-purified by flash columnchromatography (silica gel, 5% to 10% EtOAc/CH₂Cl₂ gradient) to give amixture of epoxide 142 and DIPT (1:0.85 molar ratio) as a colorless oil,which was used in the next step without additional purification. Yield(3.44 g, 85.6%); ¹H NMR (400 MHz, DMSO-d₆) δ 7.54 (t, J=1.6 Hz, 1H),7.45 (ddd, J=1.2, 2.0, 7.8 Hz, 1H), 7.34-7.38 (m, 1H), 7.29 (t, J=7.6Hz, 1H), 5.68 (d, J=4.5 Hz, 1H), 4.41 (t, J=4.7 Hz, 1H), 2.99-3.03 (m,1H), 2.69 (ABd, J=5.5, 3.9 Hz, 1H), 2.63 (ABd, J=5.3, 2.7 Hz, 1H).

Step 3.

A solution of epoxide:DIPT 142 (0.47 g, 0.803 mmol), ammonium hydroxide(25%, 5 mL) and NH₃/MeOH (7N, 5 mL) was stirred in a pressure bottle atroom temperature for 20 hrs, and then concentrated under reducedpressure. The residue was dissolved in MTBE:MeOH (1:1, 10 mL) and ethyltrifluoroacetate (3.0 mL) was added. The mixture was stirred at roomtemperature for 1 h, concentrated under reduced pressure and the residuewas purified by flash column chromatography (silica gel, 30% to 60%EtOAc/hexanes gradient) to give trifluoroacetamide 144 as a colorlessoil. Yield (0.248 g, 66%); ¹H NMR (400 MHz, DMSO-d₆) δ 9.17 (br. t, 1H),7.51 (t, J=1.8 Hz, 1H), 7.40 (ddd, J=1.2, 2.0, 7.9 Hz, 1H), 7.30-7.33(m, 1H), 7.25 (t, J=7.8 Hz, 1H), 5.57 (d, J=4.7 Hz, 1H), 4.96 (d, J=6.06Hz, 1H), 4.39 (t, J=5.5 Hz, 1H), 3.62-3.69 (m, 1H), 3.38 (dt, J=4.1,13.7 Hz, 1H), 3.05-3.13 (m, 1H).

Step 4.

Coupling of bromide 144 with olefin 120 following the method used inExample 111 except that: anhydrous degassed DMF (1 mL) was used as thereaction solvent, the reaction was heated at 90° C. for 3 hrs then at60° C. overnight. After addition of water, the product was extractedwith EtOAc (3×). To give olefin 145, yield (0.194 g, 70%); ¹H NMR (400MHz, DMSO-d₆) δ 9.14 (t, J=5.7 Hz, 1H), 7.35-7.39 (m, 1H), 7.19-7.25 (m,2H), 7.13-7.18 (m, 1H), 6.51 (d, J=16.0 Hz, 1H), 6.34 (d, J=16.0 Hz,1H), 5.41 (d, J=4.5 Hz, 1H), 4.86 (d, J=6.3 Hz, 1H), 4.39-4.43 (m, 2H),3.66-3.73 (m, 1H), 3.37 (ddd, J=3.3, 4.7, 13.3 Hz, 1H), 3.08-3.16 (m,1H), 1.55-1.67 (m, 2H), 1.37-1.54 (m, 7H), 1.18-1.25 (m, 1H).

Step 5.

A mixture of trifluoroacetamide 145 (0.189 g, 0.488 mmol), NH₃/MeOH (7N,3.0 mL) and ammonium hydroxide (10.0 mL) was stirred at room temperaturefor 68 hrs and concentrated under reduced pressure. The residue waspurified by flash chromatography using a gradient of 50% to 100% 7NNH₃/MeOH in EtOAc/hexanes to give crude amine as a colorless oil. Theamine was re-purified by flash chromatography using 20% 7N NH₃/MeOH inCH₂Cl₂ to give Example 124 as a colorless oil. Yield (0.065 g, 46%); ¹HNMR (400 MHz, MeOH-d₄) δ 7.42-7.45 (m, 1H), 7.22-7.32 (m, 3H), 6.61 (d,J=16.2 Hz, 1H), 6.36 (d, J=16.0 Hz, 1H), 4.58 (d, J=6.1 Hz, 1H),3.71-3.76 (m, 1H), 2.92 (dd, J=3.5, 13.1 Hz, 1H), 2.77 (dd, J=8.0, 13.1Hz, 1H), 1.47-1.76 (m, 9H), 1.25-1.40 (m, 1H); ¹³C NMR (100 MHz,MeOH-d₄) δ 142.7, 137.8, 137.5, 128.1, 126.9, 125.8, 125.4, 124.8, 76.1,75.7, 71.2, 43.3, 37.5, 25.5, 21.9; ESI MS m/z 292.5 [M+H]⁺; HPLC(Method 10) 97% (AUC), t_(R)=5.44 min.

Example 125 Preparation of(E)-2-(3-(3-amino-1-hydroxypropyl)styryl)cyclohexanol

(E)-2-(3-(3-amino-1-hydroxypropyl)styryl)cyclohexanol was preparedfollowing the method described in Scheme 46.

Step 1:

To an ice cold solution of 3-(3-bromophenyl)-3-hydroxypropanenitrile(119) (2.70 g, 11.9 mmol) in anhydrous THF (20 mL) under argon was addeda solution of LiAlH₄ in THF (11.9 mL of a 2 M solution in THF, 23.8mmol) The mixture was stirred at 0° C. for 45 min, diluted with ether(50 mL), and quenched with the dropwise addition of saturated aqueousNa₂SO₄ (approximately 2 mL). After drying over MgSO₄, the solution wasfiltered and concentrated under reduced pressure to give amine 146 as alight green oil. Yield (2.30 g, 84%.) This material was used in the nextstep without further purification. ¹H NMR (400 MHz, DMSO-d₆) δ 7.49 (m,1H), 7.37 (dt, J=7.2, 1.6 Hz, 1H), 7.23-7.31 (m, 2H), 4.66 (t, J=6.8 Hz,1H), 2.61 (m, 2H), 1.61 (q, J=6.8 Hz, 2H).

Step 2:

To a solution of 3-amino-1-(3-bromophenyl)propan-1-ol (146) (2.30 g, 10mmol) in anhydrous THF (20 mL) was added ethyl trifluoroacetate (4.0 mL,33.5 mmol). The reaction mixture was stirred at room temperature for 3h, then concentrated under reduced pressure. Purification by columnchromatography (10 to 70% EtOAc-hexanes gradient) gaveN-(3-(3-bromophenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamide (147) asan oil containing ˜15% of2,2,2-trifluoro-N-(3-hydroxy-3-phenylpropyl)acetamide. Yield (1.96 g,60%): ¹H NMR (400 MHz, DMSO-d₆) δ 9.33 (s, 1H), 7.51 (t, J=2.0 Hz, 1H),7.41 (dt, J=7.6, 2.0 Hz, 1H), 7.25-7.32 (m, 2H), 5.46 (d, J=6.4 Hz, 1H),4.55-4.60 (m, 1H), 3.20-3.23 (m, 2H), 1.75- 1.82 (m, 2H).

Step 3:

Heck coupling of 2-vinylcyclohexanol with bromide 147 following themethod used in Example 111 gave trifluoroamide 148 as an orange oil.Yield (0.36 mg, 66%). ¹H NMR (400 MHz, CDCl₃) δ 7.47 (brs, 1H),7.24-7.34 (m, 3H), 7.14-7.20 (m, 1H), 6.48 (d, J=16 Hz, 1H), 6.09 (q,J=8.8 Hz, 1H), 4.83 (q, J=4.4 Hz, 1H), 3.60-3.70 (m, 1H), 3.28-3.44 (m,2H), 2.38 (brs, 2H), 2.00-2.10 (m, 2H), 1.93-1.99 (m, 2H), 1.78-1.84 (m,2H), 1.64-1.73 (m, 1H), 1.22-1.40 (m, 4H).

Step 4:

Deprotection of trifluoroacetamide 148 following the method used inExample 71 gave Example 125 as a white foamy solid. Yield (0.11 g, 41%).¹H NMR (400 MHz, CDCl₃) δ 7.43 (brs, 1H), 7.18-7.31 (m, 4H), 6.52 (d,J=16 Hz, 1H), 6.06-6.13 (m, 1H), 4.91-4.97 (brs, 1H), 3.29-3.35 (m, 1H),3.04-3.16 (m, 1H), 2.90-3.00 (m, 1H), 2.65 (brs, 3H), 2.00-2.50 (2H),1.64-1.90 (m, 5H), 1.18-1.40 (m, 4H). ESI MS m/z 276.3 [m+H]⁺, 258.3[m+H−H₂O]⁺

Example 126 Preparation of(E)-1-(5-(3-amino-1-hydroxypropyl)-2-fluorostyryl)cyclohexanol

(E)-1-(5-(3-Amino-1-hydroxypropyl)-2-fluorostyryl)cyclohexanol wasprepared according to the Methods used in Examples 115.

Step 1:

Alkylation of 3-bromo-4-fluorobenzaldehyde with acetonitrile followingthe method used in Example 115 gave3-(3-bromo-4-fluorophenyl)-3-hydroxypropanenitrile as a pale yellow oil.Yield (4.2 g, 70%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.71 (dd, J=6.8, 2.0 Hz,1H), 7.44 (ddd, J=8.4, 5.2, 2.4 Hz, 1H), 7.35 (t, J=8.8 Hz, 1H), 6.08(bs, 1H), 4.90 (s, 1H), 2.90 (ABd, J=16.8, 5.2 Hz, 1H), 2.83 (ABd,J=16.8, 6.4 Hz, 1H).

Step 2:

Reduction of 3-(3-bromo-4-fluorophenyl)-3-hydroxypropanenitrile withBH₃-THF followed by protection of the amine following the method inExample 115 gaveN-(3-(3-bromo-4-fluorophenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamideas a clear oil. Yield (4.3 g, 73%): ¹H NMR (400 MHz, DMSO-d₆) δ 9.31(bs, 1H), 7.62 (dd, J=6.8, 2.0 Hz, 1H), 7.37-7.33 (m, 1H), 7.30 (t,J=8.8 Hz, 1H), 5.48 (d, J=4.4 Hz, 1H), 4.60-4.56 (m, 1H), 3.28-3.15 (m,2H), 1.84-1.71 (m, 2H).

Step 3:

Coupling of olefin 120 withN-(3-(5-bromo-4-fluorophenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamidefollowing the method used in Example 115 except the reaction was donefor 16 hr at 90° C., gave(E)-2,2,2-trifluoro-N-(3-(4-fluoro-3-(2-(1-hydroxycyclohexyl)vinyl)phenyl)-3-hydroxypropyl)acetamideas a light yellow oil. Yield (0.44 g, 51%): ¹H NMR (400 MHz, DMSO-d₆) δ9.33 (bs, 1H), 7.50 (dd, J=7.6, 2.2 Hz, 1H), 7.18 (ddd, J=8.0, 5.2, 2.0Hz, 1H), 7.08 (dd, J=10.8, 8.4 Hz, 1H), 6.65 (d, J=16.4 Hz, 1H), 6.45(d, J=16.4 Hz, 1H), 5.35 (bs, 1H), 4.56 (t, J=6.4 Hz, 1H), 4.49 (bs,1H), 3.22 (bs, 2H), 1.81-1.76 (m, 2H), 1.63-1.39 (m, 9H), 1.26-1.17 (m,1H).

Step 4:

Deprotection of(E)-2,2,2-trifluoro-N-(3-(4-fluoro-3-(2-(1-hydroxycyclohexyl)vinyl)phenyl)-3-hydroxypropyl)acetamidefollowing the method used in Example 115 gave Example 126 as an offwhite solid. Yield (0.153 g, 47%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.48 (dd,J=7.6, 2.2 Hz, 1H), 7.15 (ddd, J=7.8, 5.1, 2.2 Hz, 1H), 7.06 (dd,J=10.8, 8.4 Hz, 1H), 6.65 (d, J=16.4 Hz, 1H), 6.43 (d, J=16.4 Hz, 1H),4.64 (m, 1H), 4.49 (bs, 1H), 2.66-2.54 (m, 2H), 1.65-1.56 (m, 4H),1.53-1.40 (m, 7H), 1.27-1.17 (m, 1H).

Example 127 Preparation of(E)-1-(3-(3-amino-1-hydroxypropyl)-5-methoxystyryl)cyclohexanol

(E)-1-(3-(3-Amino-1-hydroxypropyl)-5-methoxystyryl)cyclohexanol wasprepared according to the Methods used in Examples 115

Step 1:

Alkylation of 3-bromo-5-methoxybenzaldehyde with acetonitrile followingthe method used in Example 115 gave3-(3-bromo-5-methoxyphenyl)-3-hydroxypropanenitrile as a pale yellowoil. Yield (4.1 g, 70%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.16-7.15 (m, 1H),7.04-7.03 (m, 1H), 6.97-6.96 (m, 1H), 6.04 (d, J=4.8 Hz, 1H), 4.87-4.83(m, 1H), 3.74 (s, 3H), 2.89 (ABd, J=16.4, 5.2 Hz, 1H), 2.81 (ABd,J=16.8, 6.8 Hz, 1H).

Step 2:

Reduction of 3-(3-bromo-5-methoxyphenyl)-3-hydroxypropanenitrile withBH₃-THF followed by protection of the amine following the method inExample 115 gaveN-(3-(3-bromo-5-methoxyphenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamideas a clear oil. Yield (3.9 g, 68%): ¹H NMR (400 MHz, DMSO-d₆) δ 9.30(bs, 1H), 7.07 (t, J=1.2 Hz, 1H), 6.98-6.97 (m, 1H), 6.88-6.87 (m, 1H),5.44 (d, J=4.8 Hz, 1H), 4.56-4.51 (m, 1H), 3.74 (m, 3H), 3.27-3.15 (m,2H), 1.96-1.70 (m, 2H).

Step 3:

Coupling of olefin 120 withN-(3-(5-bromo-5-methoxyphenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamidefollowing the method used in Example 115 except the reaction was donefor 16 hr at 90° C., gave(E)-2,2,2-trifluoro-N-(3-hydroxy-3-(3-(2-(1-hydroxycyclohexyl)vinyl)-5-methoxyphenyl)propyl)acetamideas a light yellow oil. Yield (0.393 g, 48%): ¹H NMR (400 MHz, DMSO-d₆) δ9.32 (t, J=5.2, 1H), 6.93 (s, 1H), 6.79-6.78 (m, 1H), 6.73 (s, 1H), 6.47(d, J=16.0 Hz, 1H), 6.34 (d, J=16.0 Hz, 1H), 5.29 (bs, 1H), 4.52 (t,=6.0 Hz, 1H), 4.39 (bs, 1H), 3.73 (s, 3H), 3.25-3.20 (m, 2H), 1.81-1.71(m, 2H), 1.62-1.40 (m, 9H), 1.25-1.17 (m, 1H).

Step 4:

Deprotection of(E)-2,2,2-trifluoro-N-(3-hydroxy-3-(3-(2-(1-hydroxycyclohexyl)vinyl)-5-methoxyphenyl)propyl)acetamidefollowing the method used in Example 115 gave Example 127 as a clearoil. Yield (0.162 g, 55%): ¹H NMR (400 MHz, DMSO-d₆) δ 6.91 (s, 1H),6.76-6.75 (m, 1H), 6.71 (s, 1H), 6.46 (d, J=16.0 Hz, 1H), 6.33 (d,J=16.0 Hz, 1H), 4.59 (t, J=6.4 Hz, 1H), 4.39 (bs, 1H), 3.72 (s, 3H),2.66-2.55 (m, 2H), 1.63-1.57 (m, 4H), 1.48-1.39 (m, 7H), 1.25-1.15 (m,1H).

Example 128 Preparation of(E)-1-(3-(3-amino-1-hydroxypropyl)-4-fluorostyryl)cyclohexanol

(E)-1-(3-(3-amino-1-hydroxypropyl)-4-fluorostyryl)cyclohexanol wasprepared according to the Methods used in Examples 123.

Step 1:

Alkylation of 5-bromo-2-fluorobenzaldehyde gave3-(5-bromo-2-fluorophenyl)-3-hydroxypropanenitrile, which was useddirectly in the next step.

Step 2: Reduction of 3-(5-bromo-2-fluorophenyl)-3-hydroxypropanenitrileusing borane methyl sulfide3-amino-1-(5-bromo-2-fluorophenyl)propan-1-ol which was used directly inthe next step.

Step 3:

Treatment of 3-amino-1-(5-bromo-2-fluorophenyl)propan-1-ol with ethyltrifluoroacetate gaveN-(3-(5-bromo-2-fluorophenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamideas a light colored oil. Yield (0.95 g, 28% in 3 steps): ¹H NMR (400 MHz,DMSO-d₆) δ 9.37 (t, J=4.8 Hz, 1H), 7.59 (dd, J=6.4, 2.8 Hz, 1H),7.44-7.47 (m, 1H), 7.12 (dd, J=10.4, 8.8 Hz, 1H), 5.57 (d, J=4.8, Hz,1H), 4.78-4.84 (m, 1H), 3.26 (q, J=6.8, Hz, 2H), 1.71-1.84 (m, 2H).

Step 4:

Coupling ofN-(3-(5-bromo-2-fluorophenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamidewith olefin 120 gave(E)-2,2,2-trifluoro-N-(3-(2-fluoro-5-(2-(1-hydroxycyclohexyl)vinyl)phenyl)-3-hydroxypropyl)acetamideas a light yellow oil. Yield (0.19 g, 42%): ¹H NMR (400 MHz, MeOD) δ7.54 (dd, J=7.2, 2.4 Hz, 1H), 7.27-7.31 (m, 1H), 6.97 (dd, J=10.4, 8.8Hz, 1H), 6.58 (d, J=16.4 Hz, 1H), 6.30 (d, J=16 Hz, 1H), 4.99 (dd,J=8.0, 4.2 Hz, 1H), 4.41 (t, J=7.2 Hz, 2H), 1.91-2.02 (m, 2H), 1.50-1.78(m, 9H), 1.28-1.39 (m, 1H).

Step 5:

Deprotection of(E)-2,2,2-trifluoro-N-(3-(2-fluoro-5-(2-(1-hydroxycyclohexyl)vinyl)phenyl)-3-hydroxypropyl)acetamidefollowed by HCL salt formation following the methods used in Example 123gave. Example 128 hydrochloride as a light color solid. Yield (0.08 g,49%): ¹H NMR (400 MHz, MeOD) δ 7.59 (dd, J=6.4, 2.4 Hz, 1H), 7.32-7.36(m, 1H), 6.99 (dd, J=10.4, 8.8 Hz, 1H), 6.74 (d, J=16.0 Hz, 1H), 6.43(d, J=16.4 Hz, 1H), 5.89 (t, J=2.4 Hz, 1H), 5.11 (dd, J=8.4, 4.4 Hz,1H), 3.02-3.16 (m, 2H), 2.14-2.38 (m, 4H), 1.95-2.08 (m, 3H), 1.54-1.78(m, 5H).

Example 129 Preparation of(1R,2S)-3-amino-1-(3-((E)-2-(1-hydroxycyclohexyl)vinyl)phenyl)propane-1,2-diol

(1R,2S)-3-amino-1-(3-((E)-2-(1-hydroxycyclohexyl)vinyl)phenyl)propane-1,2-diolis prepared according to the Methods used in Examples 124.

Step 1.

Epoxidation of (S)-1-(3-bromophenyl)prop-2-en-1-ol (141) following themethod used in Example 124 except that one equivalent of t-BuOOH andD-(−)-diisopropyl tartrate were used gave(R)-(3-bromophenyl)((S)-oxiran-2-yl)methanol as a mixture with DIPT(1:1.5 molar ratio) as a colorless oil which was used in the next stepwithout further purification. Yield (4.12 g). ¹H NMR (400 MHz, DMSO-d₆)δ 7.50 (t, J=1.6 Hz, 1H), 7.40 (ddd, J=1.2, 2.0, 7.6 Hz, 1H), 7.29-7.33(m, 1H), 7.25 (t, J=7.6 Hz, 1H), 5.52 (d, J=5.1 Hz, 1H), 5.26 (d, J=5.9Hz, 1H), 4.51 (t, J=4.7 Hz, 1H), 3.15 (dd, J=3.3, 12.5 Hz, 1H), 3.02(dd, J=8.0, 12.7 Hz, 1H).

Step 2.

Epoxide ring opening and trifluoroacetamide protection following themethod used in Example 124 affordedN-((2S,3R)-3-(3-bromophenyl)-2,3-dihydroxypropyl)-2,2,2-trifluoroacetamideas a colorless oil. Yield (0.322 g, 42%). ¹H NMR (400 MHz, DMSO-d₆) δ9.17 (br. t, 1H), 7.51 (t, J=1.8 Hz, 1H), 7.40 (ddd, J=1.2, 2.0, 7.9 Hz,1H), 7.30-7.33 (m, 1H), 7.25 (t, J=7.8 Hz, 1H), 5.57 (d, J=4.7 Hz, 1H),4.96 (d, J=6.06 Hz, 1H), 4.39 (t, J=5.5 Hz, 1H), 3.62-3.69 (m, 1H), 3.38(dt, J=4.1, 13.7 Hz, 1H), 3.05-3.13 (m, 1H).

Step 3.

Coupling ofN-((2S,3R)-3-(3-bromophenyl)-2,3-dihydroxypropyl)-2,2,2-trifluoroacetamidewith alkene 119 was performed following the method used in Example 124to giveN-((2S,3R)-2,3-dihydroxy-3-(3-((E)-2-(1-hydroxycyclohexyl)vinyl)phenyl)propyl)-2,2,2-trifluoroacetamideas a brownish foam. Yield (0.xxx g, xx %). ¹H NMR (400 MHz, DMSO-d₆) δ9.14 (t, J=5.7 Hz, 1H), 7.35-7.39 (m, 1H), 7.19-7.25 (m, 2H), 7.13-7.18(m, 1H), 6.51 (d, J=16.0 Hz, 1H), 6.34 (d, J=16.0 Hz, 1H), 5.41 (d,J=4.5 Hz, 1H), 4.86 (d, J=6.3 Hz, 1H), 4.39-4.43 (m, 2H), 3.66-3.73 (m,1H), 3.37 (ddd, J=3.3, 4.7, 13.3 Hz, 1H), 3.08-3.16 (m, 1H), 1.55-1.67(m, 2H), 1.37-1.54 (m, 7H), 1.18-1.25 (m, 1H).

Step 4.

N-((2S,3R)-2,3-dihydroxy-3-(3-((E)-2-(1-hydroxycyclohexyl)vinyl)phenyl)propyl)-2,2,2-trifluoroacetamidewas deprotected following the method used in Example 124 to give Example129 as a colorless oil. Yield (0.xx g, xx %). ¹H NMR (400 MHz, MeOH-d₄)δ 7.42-7.45 (m, 1H), 7.22-7.32 (m, 3H), 6.61 (d, J=16.2 Hz, 1H), 6.36(d, J=16.0 Hz, 1H), 4.58 (d, J=6.1 Hz, 1H), 3.71-3.76 (m, 1H), 2.92 (dd,J=3.5, 13.1 Hz, 1H), 2.77 (dd, J=8.0, 13.1 Hz, 1H), 1.47-1.76 (m, 9H),1.25-1.40 (m, 1H). ¹³C NMR (100 MHz, MeOH-d₄) δ 142.7, 137.8, 137.5,128.1, 126.9, 125.8, 125.4, 124.8, 76.1, 75.7, 71.2, 43.3, 37.5, 25.5,21.9; ESI MS m/z 292.3 [M+H]⁺; HPLC (Method 9) 99% (AUC), t_(R)=5.31min.

Example 130 Preparation of(E)-1-(3-(3-amino-1-hydroxypropyl)-5-chlorostyryl)cyclohexanol

(E)-1-(3-(3-amino-1-hydroxypropyl)-5-chlorostyryl)cyclohexanol isprepared according to the Methods used in Examples 115.

Step 1:

Alkylation of 5-bromo-3-chlorobenzaldehyde with acetonitrile followingthe method used in Example 115 gave3-(5-bromo-3-chlorophenyl)-3-hydroxypropanenitrile as a clear oil. Yield(3.21 g, 54%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.62 (t, J=2.0 Hz, 1H),7.58-7.57 (m, 1H), 7.49-4.48 (m, 1H), 6.18 (d, J=4.8 Hz, 1H), 4.93-4.90(m, 1H), 2.93 (ABd, J=16.8, 5.2 Hz, 1H), 2.86 (ABd, J=17.2, 6.8 Hz, 1H).

Step 2:

Reduction of 3-(5-bromo-3-chlorophenyl)-3-hydroxypropanenitrile withBH₃-THF followed by protection of the amine following the method inExample 115 gaveN-(3-(5-bromo-3-chlorophenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamideas a clear oil. Yield (3.15 g, 71%): ¹H NMR (400 MHz, DMSO-d₆) δ 9.30(bs, 1H), 7.55 (t, J=2.0 Hz, 1H), 7.49-7.48 (m, 1H), 7.394-7.387 (m,1H), 5.57 (d, J=4.8 Hz, 1H), 3.30-3.15 (m, 2H), 1.86-1.70 (m, 2H).

Step 3:

Coupling of olefin 119 withN-(3-(5-bromo-3-chlorophenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamidefollowing the method used in Example 115 except the reaction was donefor 16 hr at 90° C., gave(E)-N-(3-(3-chloro-5-(2-(1-hydroxycyclohexyl)vinyl)phenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamideas a clear oil. Yield (0.521 g, 58%): ¹H NMR (400 MHz, DMSO-d₆) δ 9.32(t, J=5.0 Hz, 1H), 7.319-7.310 (m, 1H), 7.30 (s, 1H), 7.194-7.187 (m,1H), 6.50 (d, J=16.0 Hz, 1H), 6.44 (d, J=16.0 Hz, 1H), 5.43 (d, J=4.8Hz, 1H), 4.59-4.54 (m, 1H), 4.44 (s, 1H), 3.26-3.16 (m, 2H), 1.86-1.72(m, 2H), 1.62-1.56 (m, 2H), 1.51-1.39 (m, 7H), 1.25-1.17 (m, 1H).

Step 4:

Deprotection of(E)-N-(3-(3-chloro-5-(2-(1-hydroxycyclohexyl)vinyl)phenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamidefollowing the method used in Example 115 gave Example 130 as clear oil.Yield (0.3 g, 76%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.283-7.275 (m, 2H),7.169-7.162 (m, 1H), 6.49 (d, J=16.0 Hz, 1H), 6.43 (d, J=16.0 Hz, 1H),4.64 (t, J=5.8 Hz, 1H), 4.44 (bs, 1H), 2.66-2.54 (m, 2H), 1.63-1.56 (m,4H), 1.51-1.39 (m, 7H), 1.25-1.63 (m, 1H).

Example 131 Preparation of(E)-4-(3-(2,6-dimethylstyryl)phenyl)butan-1-amine

(E)-4-(3-(2,6-Dimethylstyryl)phenyl)butan-1-amine was prepared followingthe method described in Scheme 47.

Step 1:

To a stirred solution of (E)-2-(3-iodostyryl)-1,3-dimethylbenzene (4)(0.143 g, 0.428 mmol), tris(dibenzylideneacetone)dipalladium(0) (0.020g, 0.022 mmol) and tri(o-tolyl)phosphine (0.026 g, 0.085 mmol) in DMF (2mL) was added 4-ethoxy-4-oxobutylzinc bromide (0.86 mL, 0.5M solution inTHF, 0.430 mmol) at room temperature. After 22 h water (20 mL) wasadded, the mixture was extracted with ethyl acetate (3×50 mL), thecombined extracts were washed with water (3×20 mL) and brine (20 mL),dried (Na₂SO₄), filtered and concentrated. The resulting residue waspurified by flash column chromatography (silica gel, 96:4 hexanes/ethylacetate) to give 149 as a colorless oil. Yield (0.119 g, 86%): ESI MSm/z 277 [M+H−EtOH]⁺.

Step 2:

To a stirred solution of 149 (0.119 g, 0.369 mmol) in methanol (5 mL),THF (5 mL) and water (3 mL) was added lithium hydroxide (0.088 g, 3.67mmol) at room temperature. After 3 h the reaction mixture wasconcentrated, the residue was diluted with brine (10 mL) and theresulting mixture was acidified with 4N hydrochloric acid to pH 2. Theresulting mixture was extracted with ethyl acetate (3×50 mL) and thecombined extracts were dried (Na₂SO₄), filtered and concentrated. Theresulting residue was purified by flash column chromatography (silicagel, 90:10 methylene chloride/methanol) to give 150 as a colorlesssyrup. Yield (0.105 g, 96%): ¹H NMR (300 MHz, CDCl₃) δ 7.43-7.07 (m,8H), 6.57 (d, J=16.6 Hz, 1H), 2.70 (t, J=7.3 Hz, 2H), 2.41 (t, J=7.3 Hz,2H), 2.37 (s, 6H), 2.00 (m, 2H); ESI MS m/z 277 [M+H−H₂O]⁺.

Step 3:

To a stirred solution of 150 (0.105 g, 0.357 mmol) in DMF (5 mL) wasadded N,N-diisopropylethylamine (0.230 g, 1.78 mmol),1-hydroxybenzotriazole (0.097 g, 0.717 mmol),1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.137 g,0.715 mmol) and ammonium chloride (0.038 g, 0.710 mmol) at roomtemperature. After 16 h the reaction mixture was diluted with ethylacetate (100 mL) and washed sequentially with 10% aqueous potassiumcarbonate (20 mL), water (2×20 mL) and brine (20 mL). The organic layerwas dried (Na₂SO₄), filtered and concentrated. The residue was purifiedby flash column chromatography (silica gel, 60:40 methylenechloride/ethyl acetate) to give 151 as a colorless oil. Yield (0.040 g,38%): ¹H NMR (500 MHz, CDCl₃) δ 7.32 (m, 3H), 7.08 (m, 5H), 6.57 (d,J=16.6 Hz, 1H), 5.60 (br s, 1H), 5.48 (br s, 1H), 2.70 (t, J=7.3 Hz,2H), 2.26 (t, J=7.3 Hz, 2H), 2.04 (s, 6H), 2.01 (m, 2H); ESI MS m/z 294[M+H]⁺.

Step 4:

To a stirred solution of 151 (0.040 g, 0.136 mmol) in THF (5 mL) wasadded lithium aluminum hydride (0.052 g, 1.37 mmol) at room temperature.After 66 h the reaction mixture was cooled to 0° C., quenched with 2Naqueous sodium hydroxide (0.1 mL), the resulting suspension was dilutedwith MTBE (50 mL), filtered and concentrated. The resulting residue waspurified by flash column chromatography (silica gel, 50:46:4 ethylacetate/hexanes/7N ammonia in methanol) followed by preparative HPLC togive Example 132 as a colorless oil. Yield (0.017 g, 45%): R_(f) 0.65(silica gel, 50:40:10 ethyl acetate/hexanes/7N ammonia in methanol); ¹HNMR (500 MHz, CD₃OD) δ 7.35 (m, 2H), 7.26 (t, J=7.7 Hz, 1H), 7.16 (d,J=16.4 Hz, 1H), 7.11 (d, J=7.5 Hz, 1H), 7.03 (s, 3H), 6.57 (d, J=16.4Hz, 1H), 2.68 (t, J=6.9 Hz, 2H), 2.66 (t, J=6.9 Hz, 2H), 2.34 (s, 6H),1.68 (m, 2H), 1.54 (m, 2H); ¹³C NMR (125 MHz, CD₃OD) δ 144.2, 139.1,138.4, 137.3, 135.6, 129.8, 128.99, 128.98, 127.9, 127.8, 127.7, 124.9,42.5, 36.8, 33.2, 30.1, 21.3; ESI MS m/z 280 [M+H]⁺; HPLC (Method E)92.6% (AUC), t_(R)=13.32 min. HRMS Calcd for C₂₀H₂₅N [M+H]: 280.2065.Found: 280.2064.

Example 132 Preparation of1-(3-(2,6-dimethylstyryl)phenyl)-N,N-dimethylmethanamine

1-(3-(2,6-Dimethylstyryl)phenyl)-N,N-dimethylmethanamine was preparedfollowing the method used in Example 4.

Step 1:

Coupling of (2,6-dimethylbenzyl)triphenylphosphonium bromide with methyl3-formylbenzoate following the method described in Example 1 gave methyl3-(2,6-dimethylstyryl)benzoate as a white solid 2.20 g (71%), isomerratio 2:1 trans: cis.

cis-isomer: ¹H NMR (500 MHz, CDCl₃) δ 7.81-7.78 (m, 1H), 7.71-7.67 (m,1H), 7.22-7.03 (m, 5H), 6.72-6.62 (m, 2H), 2.37 (s, 6H), 2.15 (s, 3H);

trans-isomer: ¹H NMR (500 MHz, CDCl₃) δ 8.15 (d, J=1.6 Hz, 1H),7.96-7.93 (m, 1H), 7.71-7.67 (m, 1H), 7.45 (t, J=7.8 Hz, 1H), 7.22-7.03(m, 4H), 6.72-6.62 (m, 1H), 2.37 (s, 6H), 2.15 (s, 3H); ESI MS m/z 267[M+H]⁺.

Step 2:

Hydrolysis of methyl 3-(2,6-dimethylstyryl)benzoate gave3-(2,6-dimethylstyryl)benzoic acid as a white solid. Yield (2.16 g,quant.), isomer ratio 2:1 trans:cis: ¹H NMR (500 MHz, CDCl₃) δ 7.87 (d,J=7.5 Hz, 1H), 7.82 (s, 1H), 7.23-7.05 (m, 5H), 6.73-6.64 (m, 2H), 2.15(s, 6H); The trans-isomer: ¹H NMR (500 MHz, CDCl₃) δ 8.24 (s, 1H), 8.02(d, J=7.7 Hz, 1H), 7.74 (d, J=7.6 Hz, 1H), 7.49 (t, J=7.7 Hz, 1H),7.22-7.03 (m, 4H), 6.73-6.64 (m, 1H), 2.38 (s, 6H).

Step 3:

Coupling of 3-(2,6-dimethylstyryl)benzoic acid with dimethylamine gave3-(2,6-Dimethylstyryl)-N,N-dimethylbenzamide as a yellow oil. Yield(0.223 g, 81%): cis-isomer: ¹H NMR (500 MHz, CDCl₃) δ 7.23-7.17 (m, 2H),7.11-6.98 (m, 5H), 6.67 (d, J=12.2 Hz, 1H), 6.58 (d, J=12.2 Hz, 1H),3.00 (s, 3H), 2.66 (s, 3H), 2.15 (s, 6H); trans- isomer: ¹H NMR (500MHz, CDCl₃) δ 7.56 (s, 1H), 7.52 (d, J=7.9 Hz, 1H), 7.39 (t, J=7.6 Hz,1H), 7.29 (d, J=7.5 Hz, 1H), 7.14 (d, J=16.6 Hz, 1H), 7.11-6.98 (m, 3H),6.59 (d, J=16.6 Hz, 1H), 3.14 (s, 3H), 3.02 (s, 3H), 2.36 (s, 6H); ESIMS m/z 280 [M+H]⁺.

Step 4:

Reduction of 3-(2,6-Dimethylstyryl)-N,N-dimethylbenzamide gave Example133 as a yellow oil. Yield (0.062 g, 29%), isomer ratio 3.3:1trans:cis:R_(f) 0.95 (silica gel, 95:5 methylene chloride/7N ammonia inmethanol); ¹H NMR (500 MHz, CD₃OD) 5 The cis-isomer: ¹H NMR (500 MHz,CDCl₃) δ 7.10-6.99 (m, 6H), 6.93-6.92 (m, 1H), 6.86 (s, 1H), 6.69 (d,J=12.1 Hz, 1H), 3.23 (s, 2H), 2.12 (s, 6H), 2.06 (s, 6H); Thetrans-isomer: ¹H NMR (500 MHz, CDCl₃) δ 7.51 (s, 1H), 7.45 (d, J=7.7 Hz,1H), 7.33 (t, J=7.7 Hz, 1H), 7.22-7.19 (m, 2H), 7.10-6.99 (m, 3H), 6.59(d, J=16.6 Hz, 1H), 3.50 (s, 2H), 2.34 (s, 6H), 2.26 (s, 6H); ¹³C NMR(75 MHz, CD₃OD) δ 139.2, 139.1, 138.9, 138.3, 137.1, 136.5, 135.1,132.1, 130.7, 130.2, 129.9, 129.7, 129.3, 128.9, 128.6, 128.5, 128.4,128.3, 128.0, 127.8, 126.6, 64.9, 64.7, 45.3, 44.9, 21.2, 20.4; ESI MSm/z 266 [M+H]⁺; HPLC (Method E) 98.9% (AUC), t_(R)=12.55 min. HRMS calcdfor C₁₉H₂₃N [M+H]: 266.1908. Found: 266.1903.

Example 133 Preparation of 4-(3-(2,6-dimethylstyryl)benzyl)morpholine

4-(3-(2,6-Dimethylstyryl)benzyl)morpholine was prepared following themethod used in Example 4.

Step 1:

Coupling of 3-(2,6-dimethylstyryl)benzoic acid with N-methyl morpholinegave (3-(2,6-Dimethylstyryl)phenyl)(morpholino)methanone as a colorlessoil. Yield (0.344 g, quant.) isomer ratio 2:1 trans:cis: cis-isomer: ¹HNMR (500 MHz, CDCl₃) δ 7.28-7.22 (m, 2H), 7.19-7.06 (m, 4H), 6.93 (s,1H), 6.68 (d, J=12.3 Hz, 1H), 6.60 (d, J=12.2 Hz, 1H), 3.79-3.50 (m,8H), 2.14 (s, 6H); trans-isomer: ¹H NMR (500 MHz, CDCl₃) δ 7.54 (d,J=8.7 Hz, 2H), 7.40 (d, J=7.6 Hz, 1H), 7.28-7.22 (m, 1H), 7.15 (d,J=16.7 Hz, 1H), 7.19-7.06 (m, 2H), 7.03 (d, J=7.6 Hz, 1H), 6.61 (d,J=16.7 Hz, 1H), 3.79-3.50 (m, 8H), 2.36 (s, 6H); ESI MS m/z 322 [M+H]⁺.

Step 2:

Reduction of (3-(2,6-Dimethylstyryl)phenyl)(morpholino)methanone gaveExample 134 as a yellow oil. Yield (0.030 g, 10%), isomer ration 3:1trans:cis::R_(f) 0.66 (silica gel, ethyl acetate); cis-isomer: ¹H NMR(500 MHz, CDCl₃) δ 7.26-7.24 (m, 1H), 7.10-7.08 (m, 4H), 6.91-6.89 (m,2H), 6.65 (d, J=12.3 Hz, 1H), 6.53 (d, J=12.3 Hz, 1H), 3.64 (t, J=4.7Hz, 4H), 3.30 (s, 2H), 2.26 (t, J=4.3 Hz, 4H), 2.14 (s, 6H);trans-isomer: ¹H NMR (500 MHz, CDCl₃) δ 7.43 (d, J=6.4 Hz, 2H), 7.33 (d,J=7.8 Hz, 1H), 7.26-7.24 (m, 1H), 7.11 (d, J=16.7 Hz, 1H), 7.10-7.08 (m,2H), 7.02 (d, J=7.5 Hz, 1H), 6.59 (d, J=16.7 Hz, 1H), 3.73 (t, J=4.7 Hz,4H), 3.53 (s, 2H), 2.49 (d, J=3.8 Hz, 4H), 2.37 (s, 6H); ¹³C NMR (75MHz, CDCl₃) δ 167.2, 138.0, 137.4, 137.0, 136.6, 136.2, 135.7, 135.3,134.5, 133.3, 130.8, 130.3, 129.9, 128.9, 128.8, 128.6, 128.1, 127.9,127.6, 127.1, 126.9, 125.9, 125.6, 125.1, 53.4, 41.0, 40.4, 39.7, 38.8,32.0, 31.4, 21.1, 20.2; ESI MS m/z 308 [M+H]⁺; HPLC (Method E) 97.2%(AUC), t_(R)=14.7 min. HRMS calcd for C₂₁H₂₅NO [M+H]: 308.2014. Found:308.2003.

Example 134 Preparation of(E)-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)methanamine

(E)-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)methanamine wasprepared following the method used in Example 3 and 32.

Step 1:

Coupling of Wittig salt 3 with methyl 3-formyl benzoate following themethod used in Example 32 gave methyl3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)benzoate (0.698 g, 88%) as awhite foam: ¹H NMR (500 MHz, CDCl₃) δ 8.14 (s, 1H), 7.96 (d, J=7.7 Hz,1H), 7.64 (d, J=7.7 Hz, 1H), 7.42 (m, 1H), 6.77 (d, J=16.3 Hz, 1H), 6.39(d, J=16.3 Hz, 1H), 2.05 (t, J=6.1 Hz, 2H), 1.77 (s, 3H), 1.65 (m, 2H),1.50 (m, 2H), 1.08 (s, 6H); ESI MS m/z 271 [M+H]⁺.

Step 2:

Hydrolysis of methyl 3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)benzoatefollowing the procedure used in Example 3 gave3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)benzoic acid as a white foam.Yield (0.698 g, 88%): ¹H NMR (500 MHz, CDCl₃) δ 8.14 (s, 1H), 7.96 (d,J=7.7 Hz, 1H), 7.64 (d, J=7.7 Hz, 1H), 7.42 (m, 1H), 6.77 (d, J=16.3 Hz,1H), 6.39 (d, J=16.3 Hz, 1H), 2.05 (t, J=6.1 Hz, 2H), 1.77 (s, 3H), 1.65(m, 2H), 1.50 (m, 2H), 1.08 (s, 6H); ESI MS m/z 271 [M+H]⁺.

Step 3:

Amidation of 3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)benzoic acidfollowing the procedure used in Example 3 gave3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)benzamide as a white foam.Yield (0.279 g, 40%): ¹H NMR (500 MHz, CDCl₃) δ 7.86 (t, J=1.5 Hz, 1H),7.61 (d, J=7.7 Hz, 1H), 7.56 (d, J=7.7 Hz, 1H), 7.39 (t, J=7.7 Hz, 1H),6.75 (d, J=16.3 Hz, 1H), 6.37 (d, J=16.3 Hz, 1H), 6.09 (br s, 1H), 5.71(br s, 1H), 2.04 (t, J=6.1 Hz, 2H), 1.76 (s, 3H), 1.65 (m, 2H), 1.48 (m,2H), 1.06 (s, 6H); ESI MS m/z 270 [M+H]⁺.

Step 4:

Reduction of 3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)benzamidefollowing the procedure used in Example 3 gave Example 135 as a lightyellow oil. Yield (0.135 g, 51: R_(f) 0.45 (silica gel, 50:35:15hexanes/ethyl acetate/7N ammonia in methanol); ¹H NMR (500 MHz, CD₃OD) δ7.40 (s, 1H), 7.27 (m, 2H), 7.17 (m, 1H), 6.73 (d, J=16.3 Hz, 1H), 6.33(d, J=16.3 Hz, 1H), 3.78 (s, 2H), 2.05 (t, J=6.0 Hz, 2H), 1.75 (s, 3H),1.67 (m, 2H), 1.51 (m, 2H), 1.06 (s, 6H); ¹³C NMR (125 MHz, CD₃OD) δ143.8, 139.6, 139.0, 134.4, 130.4, 129.8, 128.7, 127.3, 126.1, 125.7,46.7, 40.8, 35.3, 33.9, 29.4, 21.9, 20.4; ESI MS m/z 256 [M+H]⁺; HPLC(Method E) 98.2% (AUC), t_(R)=11.54 min. HRMS Calcd for C₁₈H₂₅N[M+H−NH₃]: 239.1800. Found: 239.1793.

Example 135 Preparation of(E)-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)methanamine

(E)-(3-(2-(2,6,6-trimethylcyclohex-1-enyl)vinyl)phenyl)methanamine wasprepared following the method used in Example 1.

Step 1:

Amidation of aldehyde 69 with ethanolamine following the method used inExample 1 gave Example 136 as a mixture of trans- and cis-isomers.Further purification by preparative HPLC provided Example 136 as acolorless oil. Yield (0.063 g, 37%): R_(f) 0.27 (silica gel, 90:10Methylene Chloride/7N Ammonia in Methanol); ¹H NMR (500 MHz, CD₃OD) δ7.55 (s, 1H), 7.44 (d, J=7.5 Hz, 1H), 7.33 (d, J=7.6 Hz, 1H), 7.25 (d,J=7.5 Hz, 1H), 7.21 (d, J=16.6 Hz, 1H), 7.03 (s, 3H), 6.60 (d, J=16.7Hz, 1H), 3.82 (s, 2H), 3.69 (t, J=5.6 Hz, 2H), 2.76 (t, J=5.6 Hz, 2H),2.34 (s, 6H); ESI MS m/z 282 [M+H]⁺; HPLC (Method B) 91.0% (AUC),t_(R)=7.66 min. HRMS calcd for C₁₉H₂₃NO [M+H]: 282.1858. Found:282.1846.

Example 136 (E)-2-(3-(2,6-dimethylstyryl)phenyl)ethanamine

(E)-2-(3-(2,6-dimethylstyryl)phenyl)ethanamine was prepared according tothe methods used in Examples 1, 45, 50 and 55.

Step 1:

Protection of 2-(3-bromophenyl)ethanamine following the method used inExample 55 gave tert-butyl 3-bromophenethylcarbamate as a colorless oil.Yield (6.75 g, 100%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.35-7.38 (m, 2H),7.17-7.25 (m, 2H), 6.84 (t, J=5.2 Hz, 1H), 3.08-3.16 (m, 2H), 2.67 (t,J=7.2 Hz, 2H), 1.33 (s, 9H).

Step 2:

Conversion of tert-butyl 3-bromophenethylcarbamate into tert-butyl3-formylphenethylcarbamate following the method used in Example 50 gavethe product as colorless oil. Yield (0.48, 38%). ¹H NMR (400 MHz,DMSO-d₆) δ 9.97 (s, 1H), 7.71-7.74 (m, 2H), 7.49-7.54 (m, 2H), 6.88 (t,J=5.2 Hz, 1H), 3.14-3.19 (m, 2H), 2.77 (t, J=7.2 Hz, 2H), 1.33 (s, 9H).

Step 3:

Coupling of tert-butyl 3-formylphenethylcarbamate with Wittig salt 3following the method used in Example 45 gave (E)-tert-butyl3-(2,6-dimethylstyryl)phenethylcarbamate as a light yellow oil. Yield(0.43 g, 72%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.38-7.43 (m, 2H), 7.28 (t,J=8.0 Hz, 1H), 7.18 (d, J=16.8 Hz, 1H), 7.08-7.11 (m, 1H), 6.84-7.05 (m,3H), 6.62 (d, J=16.8 Hz, 1H), 3.13-3.19 (m, 2H), 2.70 (t, J=7.2 Hz, 2H),2.31 (s, 6H), 1.34 (s, 9H).

Step 4:

Deprotection of (E)-tert-butyl 3-(2,6-dimethylstyryl)phenethylcarbamatefollowing the method used in Example 45 gave(E)-2-(3-(2,6-dimethylstyryl)phenyl)ethanamine in the form of HCl saltas white solid. Yield (0.27 g, 69%): ¹H NMR (400 MHz, DMSO-d₆) δ 7.94(bs, 3H), 7.46-7.49 (m, 2H), 7.33 (t, J=7.8 Hz, 1H), 7.22 (d, J=16.8 Hz,1H), 7.17 (d, J=7.6 Hz, 1H), 7.07 (m, 3H), 6.63 (d, J=16.8 Hz, 1H),3.01-3.12 (m, 2H), 2.87-2.91 (m, 2H), 2.31 (s, 6H).

Example 137 In Vitro Isomerase Inhibition

The capability of styrenyl derivative compounds to inhibit the activityof a visual cycle isomerase was determined.

Isomerase inhibition reactions was performed essentially as described(Stecher et al., J Biol Chem 274:8577-85 (1999); see also Golczak etal., supra). Bovine Retinal Pigment Epithelium (RPE) microsome membraneswere the source of a visual cycle isomerase.

RPE Microsome Membrane Preparation

RPE microsome membrane extracts may be purchased or prepared accordingto methods practiced in the art and stored at −80° C. Crude RPEmicrosome extracts were thawed in a 37° C. water bath, and thenimmediately placed on ice. 50 ml crude RPE microsomes were placed into a50 ml Teflon-glass homogenizer (Fisher Scientific, catalog no. 0841416M)on ice, powered by a hand-held De Walt drill, and homogenized ten timesup and down on ice under maximum speed. This process was repeated untilthe crude RPE microsome solution was homogenized. The homogenate wasthen subjected to centrifugation (50.2 Ti rotor (Beckman, Fullerton,Calif.), 13,000 RPM; 15360 Rcf) for 15 minutes at 4° C. The supernatantwas collected and subjected to centrifugation a5 42,000 RPM (160,000Rcf; 50.2 Ti rotor) for 1 hour at 4° C. The supernatant was removed, andthe pellets were suspended in 12 ml (final volume) cold 10 mM MOPSbuffer, pH 7.0. The resuspended RPE membranes in 5 ml aliquots werehomogenized in a glass-to-glass homogenizer (Fisher Scientific, catalogno. K885500-0021) to high homogeneity. Protein concentration wasquantified using the BCA protein assay according to the manufacturer'sprotocol (Pierce, Rockford, Ill.; catalog no. 23227). The homogenizedRPE preparations were stored at −80° C.

Isolation of Human Apo Cellular Retinaldehyde-Binding Protein (CRALBP)

Recombinant human apo cellular retinaldehyde-binding protein (CRALBP)was cloned and expressed according to standard methods in the molecularbiology art (see Crabb et al., Protein Science 7:746-57 (1998); Crabb etal., J Biol. Chem. 263:18688-92 (1988)). Briefly, total RNA was preparedfrom confluent ARPE19 cells (American Type Culture Collection, Manassas,Va.), cDNA was synthesized using an oligo(dT)₁₂₋₁₈ primer, and then DNAencoding CRALBP was amplified by two sequential polymerase chainreactions (see Crabb et al., J Biol. Chem. 263:18688-92 (1988); Intres,et al., J Biol. Chem. 269:25411-18 (1994); GenBank Accession No.L34219.1). The PCR product was sub-cloned into pTrcHis2-TOPO TA vectoraccording to the manufacturer's protocol (Invitrogen Inc., Carlsbad,Calif.; catalog no. K4400-01), and then the sequence was confirmedaccording to standard nucleotide sequencing techniques. Recombinant6xHis-tagged human CRALBP was expressed in One Shot TOP 10 chemicallycompetent E. coli cells (Invitrogen), and the recombinant polypeptidewas isolated from E. coli cell lysates by nickel affinity chromatographyusing Ni Sepharose XK16-20 columns for HPLC (Amersham Bioscience,Pittsburgh, Pa.; catalog no. 17-5268-02). The purified 6xHis-taggedhuman CRALBP was dialyzed against 10 mM bis-tris-Propane (BTP) andanalyzed by SDS-PAGE. The molecular weight of the recombinant humanCRALBP was approximately 39 kDal.

Isomerase Assay

Each styrenyl derivative compound and control compounds werereconstituted in ethanol to 0.1 M. Ten-fold serial dilutions (10⁻²,10⁻³, 10⁻⁴, 10⁻⁵, 10⁻⁶ M) in ethanol of each compound were prepared foranalysis in the isomerase assay.

The isomerase assay was performed in 10 mM bis-tris-propane (BTP)buffer, pH 7.5, 0.5% BSA (diluted in BTP buffer), 1 mM sodiumpyrophosphate, 20 μM all-trans retinol (in ethanol), and 6 μMapo-CRALBP. The test compounds (2 μl) (final 1/15 dilution of serialdilution stocks) were added to the above reaction mixture to which RPEmicrosomes were added. The same volume of ethanol was added to thecontrol reaction (absence of test compound). Bovine RPE microsomes (9μl) (see above) were then added, and the mixtures transferred to 37° C.to initiate the reaction (total volume=150 μl). The reactions werestopped after 30 minutes by adding methanol (300 μl). Heptane was added(300 μl) and mixed into the reaction mixture by pipetting. Retinoid wasextracted by agitating the reaction mixtures, followed by centrifugationin a microcentrifuge. The upper organic phase was transferred to HPLCvials and then analyzed by HPLC using an Agilent 1100 HPLC system withnormal phase column: SILICA (Agilent Technologies, dp 5μ, 4.6 mmX, 25CM; running method had flow rate of 1.5 ml/min; injection volume 100μl). The solvent components were 20% of 2% isopropanol in ethyl acetateand 80% of 100% hexane. The area under the A₃₁₈ nm curve represented the11-cis retinol peak, which was calculated by Agilent Chemstationsoftware and recorded manually. The IC₅₀ values (concentration ofcompound that gives 50% inhibition of 11-cis retinol formation in vitro)were calculated using GraphPad Prism® 4 Software (Irvine, Calif.). Eachexperiment was performed three times in duplicate. Inhibition ofisomerase activity (IC₅₀) was determined for each compound.

The concentration dependent effect of the compounds disclosed herein onthe retinol isomerization reaction can also be evaluated with arecombinant human enzyme system. In particular, the in vitro isomeraseassay was performed essentially as in Golczak et al. 2005, PNAS 102:8162-8167, ref. 3). A homogenate of HEK293 cell clone expressingrecombinant human RPE65 and LRAT were the source of the visual enzymes,and exogenous all-trans-retinol (about 20 μM) was used as the substrate.Recombinant human CRALBP (about 80 ug/mL) was added to enhance theformation of 11 cis-retinal. The 200 μL Bis-Tris Phosphate buffer (10mM, pH 7.2) based reaction mixture also contains 0.5% BSA, and 1 mMNaPPi. In this assay, the reaction was carried out at 37° C. induplicates for one hour and was terminated by addition of 300 μLmethanol. The amount of reaction product, 11-cis-retinol, was measuredby HPLC analysis following Heptane extraction of the reaction mixture.The Peak Area Units (PAUs) corresponding to 11 cis-retinol in the HPLCchromatograms were recorded and concentration dependent curves analyzedby GraphPad Prism for IC₅₀ values. The ability of the numerous compoundsdisclosed herein to inhibit isomerization reaction is quantified and therespective IC50 value is determined. The table below summarises the 1050values of various compounds of the present invention determined byeither of the above two methods.

TABLE 13 IC₅₀ Compound/Example Number ≤0.01 μM 111, 113, 116  ≤0.1 μM114, 115, 117, 120   ≤1 μM 1, 21, 22, 23, 24, 25, 26, 27, 31, 32, 41,43, 45, 48, 49, 50, 54, 55, 56, 59, 61, 66, 68, 72, 73, 74, 75, 79, 80,82, 83, 84, 85, 100, 105, 106   ≤10 μM 7, 16, 18, 19, 20, 28, 30, 33,34, 35, 36, 37, 39, 44, 46, 47, 51, 63, 64, 65, 67, 69, 70, 77, 78, 81,89, 93, 95, 96, 102   ≤50 μM 2, 3, 6, 8, 9, 11, 12, 15, 17, 29, 38, 40,53, 58, 60, 62, 76, 107, 108, 109  ≤100 μM 5, 10, 42, 71  ≤500 μM 4, 13,14, 57

Example 138 In Vivo Murine Isomerase Assay

The capability of styrenyl derivatives to inhibit isomerase isdetermined by an in vivo murine isomerase assay. Brief exposure of theeye to intense light (“photobleaching” of the visual pigment or simply“bleaching”) is known to photo-isomerize almost all 11-cis-retinal inthe retina. The recovery of 11-cis-retinal after bleaching can be usedto estimate the activity of isomerase in vivo. Procedures were performedessentially as described by Golczak et al., Proc. Natl. Acad. Sci. USA102:8162-67 (2005). See also Deigner et al., Science, 244: 968-71(1989); Gollapalli et al., Biochim Biophys Acta. 1651: 93-101 (2003);Parish, et al., Proc. Natl. Acad. Sci. USA, 14609-13 (1998); Radu, etal., Proc Natl Acad Sci USA 101: 5928-33 (2004). The styrenyl compoundsof the present invention are expected to inhibit an isomerase reactionresulting in production of 11-cis retinol, which reaction occurs in RPE,The expected ED50 value can be 1 mg/kg or less when measured 2 hours, 4hours, 6 hours, 8 hours or longer aftering administering to the subjectcompounds a subject compound.

Six-week old dark-adapted CD-1 (albino) male mice were orally gavagedwith compounds (0.1-25 mg/kg) dissolved in 100 μl corn oil containing10% ethanol (five animals per group). Mice were gavaged with styrenylderivative compounds, Compound 1 or Compound 25. Compound B was includedas a positive control, and Compound A with low activity was alsoincluded. After 1-72 hours in the dark, the mice were exposed tophotobleaching of 3,000 lux of white light for 10 minutes. The mice wereallowed to recover 2 hours in the dark. The animals were then sacrificedby carbon dioxide inhalation. Retinoids were extracted from the eye andthe regeneration of 11-cis-retinal was assessed at various timeintervals.

Eye Retinoid Extraction

All steps were performed in darkness with minimal redlight illumination(low light darkroom lights and redfiltered flashlights for spotillumination as needed) (see, e.g., Maeda et al., J. Neurochem85:944-956, 2003; Van Hooser et al., J Biol Chem 277:19173-82, 2002).After the mice were sacrificed, the eyes were immediately removed andplaced in liquid nitrogen. The eyes were then homogenized in aglass/glass homogenizer (Kontes Glass Co., homogenizer & pestle 21)containing 1 ml retinoid analysis buffer (50 mM MOPS, 10 mM NH₂OH, 50%EtOH, pH 7.0 (stock buffer was stored in absence of ethanol)). The eyeswere homogenized until no visible tissue remained (approximately 3minutes). The samples were incubated 20 minutes at room temperature(including homogenizing) and then placed on ice. One ml cold EtOH wasadded to the homogenate to rinse the pestle, and the homogenate mixturewas transferred to 7 ml glass screwtop tubes on ice. The homogenizer wasrinsed with 7 ml hexane, which was added to the 7 ml tubes on ice.

The homogenate was mixed by vortexing at high speed for 1 minute. Thephases were separated by centrifugation (5 minutes at 4000 rpm, 4° C.).The upper phase was collected and transferred to a clean glass testtube, taking care to avoid disturbing the interface by leavingapproximately 0.2 ml of upper phase in the tube. The tubes with thecollected upper phase were placed in a heating block at 25° C. and driedunder a stream of Argon (˜30 minutes). The lower phase was againextracted by adding 4 ml hexane, vortexing, and separating the phases bycentrifugation. The upper phase was collected as described above andpooled into the drying tubes. The dried samples were solubilized in 300μl Hexane (Fisher Optima grade) and vortexed lightly. The samples weretransferred to clean 300 μl glass inserts in HPLC vials using glasspipette and the vials were crimped shut tightly.

The samples were analyzed by HPLC (HP 1100 series or Agilent 1100series, Agilent Technologies) on a Beckman Ultraspere Si column (5μparticle diameter, 4.6 mm ID×25 cm length; Part #235341). Run parametersare as follows: flow: 1.4 ml/minute, 10% Ethylacetate+90% Hexane (cleanwith 50% ethylacetate for 20 minutes); volume per injection: 100 μl(blank sample of 300 μl was run first to equilibrate column); detectionat 360 nm (detecting 11-cis retinal oxime)). The area under the curvefor 11-cis retinal oxime was calculated by Agilent Chemstation softwareand was recorded manually. Data processing was performed using Prizmsoftware.

Positive control mice (no compound administered) were sacrificed fullydark-adapted and the eye retinoids analyzed. Light (bleached) controlmice (no compound administered) were sacrificed and retinoids isolatedand analyzed immediately after light treatment.

The isomerase inhibitory activity of Compound A and Compound 1 over timeis presented in FIG. 1. The maximum inhibitory effect was observed about16 or 6 hours after gavage with Compound A or Compound 1, respectively.FIG. 2 presents the concentration dependent inhibition of isomeraseactivity by Compound 1, Compound 25, Compound A (prodrug ACU#3223), andCompound B (ACU#3222). The estimated ED₅₀s (dose of compound that gives50% inhibition of 11-cis retinal (oxime) recovery) calculated were 9.3,4.3, 0.44 and 0.15 mg/kg for Compound A (prodrug ACU#3223), Compound1(ACU#3364), Compound 25(ACU#4178) and Compound B (ACU#3222),respectively.

Inhibition (%) 5 mg/kg 1 mg/Kg Example 2 h 4 h 6 h 24 h 2 h 4 h 6 h 24 h1 57 58 0 0 0 25 89 96 43 79 54 0 7 16 55 75 89 0 56 26 75 77 31 53 46 02 0 59 21 7 0 1 61 12 0 0 0 49 60 84 10 0 74 83 91 56 19 40 9 75 81 9085 19 20 13 79 24 0 83 97 3 55 0 111 90 0 10 mg/kg Example 2 h 4 h 6 h24 h 26 80 43 94 27 11 32 52 68 75 41 76 40 2 24 16 23 0 25 100 22 0 736 8 1 80 21 80

Negative numbers have been leveled to 0%. Some are quite low (−18%)

4231 have data at 0.1, 0.3 and 1 mg/kg

4230 have data at 10 and 25 mg/kg

4226 is 0 at 48 both doses

4203 is 44%, 5 mg/kg, 48 h

4195 is 80% 10 mg/kg, 4 h

4189 0@48 h, 5 mg/kg also 94%, 4 h@10 mg/kg

4178 have lots of data 0.1, 0.3, 1 mg/kg at very extended timepoints

4128 8% 2 48 h, 10 mg/kg4110

3364 lots more data

Example 139 Preparation of Retinal Neuronal Cell Culture System

This Example describes methods for preparing a long-term culture ofretinal neuronal cells.

All compounds and reagents are obtained from Sigma Aldrich ChemicalCorporation (St. Louis, Mo.) except as noted.

Retinal Neuronal Cell Culture

Porcine eyes are obtained from Kapowsin Meats, Inc. (Graham, Wash.).Eyes are enucleated, and muscle and tissue are cleaned away from theorbit. Eyes are cut in half along their equator and the neural retina isdissected from the anterior part of the eye in buffered saline solution,according to standard methods known in the art. Briefly, the retina,ciliary body, and vitreous are dissected away from the anterior half ofthe eye in one piece, and the retina is gently detached from the clearvitreous. Each retina is dissociated with papain (WorthingtonBiochemical Corporation, Lakewood, N.J.), followed by inactivation withfetal bovine serum (FBS) and addition of 134 Kunitz units/ml of DNaseI.The enzymatically dissociated cells are triturated and collected bycentrifugation, resuspended in Dulbecco's modified Eagle's medium(DMEM)/F12 medium (Gibco BRL, Invitrogen Life Technologies, Carlsbad,Calif.) containing 25 μg/ml of insulin, 100 μg/ml of transferrin, 60 μMputrescine, 30 nM selenium, 20 nM progesterone, 100 U/ml of penicillin,100 μg/ml of streptomycin, 0.05 M Hepes, and 10% FBS. Dissociatedprimary retinal cells are plated onto Poly-D-lysine- and Matrigel- (BD,Franklin Lakes, N.J.) coated glass coverslips that are placed in 24-welltissue culture plates (Falcon Tissue Culture Plates, Fisher Scientific,Pittsburgh, Pa.). Cells are maintained in culture for 5 days to onemonth in 0.5 ml of media (as above, except with only 1% FBS) at 37° C.and 5% CO₂.

Immunocytochemistry Analysis

The retinal neuronal cells are cultured for 1, 3, 6, and 8 weeks, andthe cells are analyzed by immunohistochemistry at each time pointImmunocytochemistry analysis is performed according to standardtechniques known in the art. Rod photoreceptors are identified bylabeling with a rhodopsin-specific antibody (mouse monoclonal, diluted1:500; Chemicon, Temecula, Calif.). An antibody to mid-weightneurofilament (NFM rabbit polyclonal, diluted 1:10,000, Chemicon) isused to identify ganglion cells; an antibody to β3-tubulin (G7121 mousemonoclonal, diluted 1:1000, Promega, Madison, Wis.) is used to generallyidentify interneurons and ganglion cells, and antibodies to calbindin(AB1778 rabbit polyclonal, diluted 1:250, Chemicon) and calretinin(AB5054 rabbit polyclonal, diluted 1:5000, Chemicon) are used toidentify subpopulations of calbindin- and calretinin-expressinginterneurons in the inner nuclear layer. Briefly, the retinal cellcultures are fixed with 4% paraformaldehyde (Polysciences, Inc,Warrington, Pa.) and/or ethanol, rinsed in Dulbecco's phosphate bufferedsaline (DPBS), and incubated with primary antibody for 1 hour at 37° C.The cells are then rinsed with DPBS, incubated with a secondary antibody(Alexa 488- or Alexa 568-conjugated secondary antibodies (MolecularProbes, Eugene, Oreg.)), and rinsed with DPBS. Nuclei are stained with4′,6-diamidino-2-phenylindole (DAPI, Molecular Probes), and the culturesare rinsed with DPBS before removing the glass coverslips and mountingthem with Fluoromount-G (Southern Biotech, Birmingham, Ala.) on glassslides for viewing and analysis.

Survival of mature retinal neurons after varying times in culture isindicated by the histochemical analyses. Photoreceptor cells areidentified using a rhodopsin antibody; ganglion cells are identifiedusing an NFM antibody; and amacrine and horizontal cells are identifiedby staining with an antibody specific for calretinin.

Cultures are analyzed by counting rhodopsin-labeled photoreceptors andNFM-labeled ganglion cells using an Olympus IX81 or CZX41 microscope(Olympus, Tokyo, Japan). Twenty fields of view are counted per coverslipwith a 20× objective lens. Six coverslips are analyzed by this methodfor each condition in each experiment. Cells that are not exposed to anystressor are counted, and cells exposed to a stressor are normalized tothe number of cells in the control.

Example 140 Effect of Styrenyl Derivative Compounds on Retinal CellSurvival

This Example describes the use of the mature retinal cell culture systemthat comprises a cell stressor for determining the effects of a styrenylderivative compound on the viability of the retinal cells.

Retinal cell cultures are prepared as described in Example 112. A2E isadded as a retinal cell stressor. After culturing the cells for 1 week,a chemical stress, A2E, is applied. A2E is diluted in ethanol and addedto the retinal cell cultures at concentration of 0, 10 μM, 20 μM, and 40μM. Cultures are treated for 24 and 48 hours. A2E is obtained from Dr.Koji Nakanishi (Columbia University, New York City, N.Y.) or issynthesized according to the method of Parish et al. (Proc. Natl. Acad.Sci. USA 95:14602-13 (1998)). A styrenyl derivative compound is thenadded to the culture. To other retinal cell cultures, a styrenylderivative compound is added before application of the stressor or isadded at the same time that A2E is added to the retinal cell culture.The cultures are maintained in tissue culture incubators for theduration of the stress at 37° C. and 5% CO₂. The cells are then analyzedby immunocytochemistry as described in Example 112.

Apoptosis Analysis

Retinal cell cultures are prepared as described in Example 1 andcultured for 2 weeks and then exposed to white light stress at 6000 luxfor 24 hours followed by a 13-hour rest period. A device was built touniformly deliver light of specified wavelengths to specified wells ofthe 24-well plates. The device contained a fluorescent cool white bulb(GE P/N FC12T9/CW) wired to an AC power supply. The bulb is mountedinside a standard tissue culture incubator. White light stress isapplied by placing plates of cells directly underneath the fluorescentbulb. The CO₂ levels are maintained at 5%, and the temperature at thecell plate is maintained at 37° C. The temperature was monitored byusing thin thermocouples. The light intensities for all devices weremeasured and adjusted using a light meter from Extech InstrumentsCorporation (P/N 401025; Waltham, Mass.). A styrenyl derivative compoundis added to wells of the culture plates prior to exposure of the cellsto white light and is added to other wells of the cultures afterexposure to white light. To assess apoptosis, TUNEL is performed asdescribed herein.

Apoptosis analysis is also performed after exposing retinal cells toblue light. Retinal cell cultures are cultured as described in Example112. After culturing the cells for 1 week, a blue light stress isapplied. Blue light is delivered by a custom-built light-source, whichconsists of two arrays of 24 (4×6) blue light-emitting diodes (SunbriteLED P/N SSP-01TWB7UWB12), designed such that each LED is registered to asingle well of a 24 well disposable plate. The first array is placed ontop of a 24 well plate full of cells, while the second one is placedunderneath the plate of cells, allowing both arrays to provide a lightstress to the plate of cells simultaneously. The entire apparatus isplaced inside a standard tissue culture incubator. The CO₂ levels aremaintained at 5%, and the temperature at the cell plate is maintained at37° C. The temperature is monitored with thin thermocouples. Current toeach LED is controlled individually by a separate potentiometer,allowing a uniform light output for all LEDs. Cell plates are exposed to2000 lux of blue light stress for either 2 hours or 48 hours, followedby a 14-hour rest period. A styrenyl derivative compound is added towells of the culture plates prior to exposure of the cells to blue lightand is added to other wells of the cultures after exposure to bluelight. To assess apoptosis, TUNEL is performed as described herein.

To assess apoptosis, TUNEL is performed according to standard techniquespracticed in the art and according to the manufacturer's instructions.Briefly, the retinal cell cultures are first fixed with 4%paraformaldehyde and then ethanol, and then rinsed in DPBS. The fixedcells are incubated with TdT enzyme (0.2 units/μl final concentration)in reaction buffer (Fermentas, Hanover, Md.) combined with Chroma-TideAlexa568-5-dUTP (0.1 μM final concentration) (Molecular Probes) for 1hour at 37° C. Cultures are rinsed with DPBS and incubated with primaryantibody either overnight at 4° C. or for 1 hour at 37° C. The cells arethen rinsed with DPBS, incubated with Alexa 488-conjugated secondaryantibodies, and rinsed with DPBS. Nuclei are stained with DAPI, and thecultures are rinsed with DPBS before removing the glass coverslips andmounting them with Fluoromount-G on glass slides for viewing andanalysis.

Cultures are analyzed by counting TUNEL-labeled nuclei using an OlympusIX81 or CZX41 microscope (Olympus, Tokyo, Japan). Twenty fields of vieware counted per coverslip with a 20× objective lens. Six coverslips areanalyzed by this method for each condition. Cells that are not exposedto a styrenyl derivative compound are counted, and cells exposed to theantibody are normalized to the number of cells in the control. Data areanalyzed using the unpaired Student's t-test. One or more subjectcompounds are expected to reduce apoptosis of retinal cells.

Example 141 Level of Rhodopsin in Animals Treated with a StyrenylDerivative Compound

This example describes determining the effect of a styrenyl derivativecompound that is a visual cycle modulator on the level of rhodopsin inthe eyes of mice after oral dosing of the animals with the compound. Thelevel of rhodopsin in the eyes is determined 6 hours after administeringthe compound to the animals.

Groups of five eight-week old male mice (20-26 grams) (strain C57/B16,Balb/c, or CD1, Charles River Laboratories, Wilmington, Mass.) arehoused at room temperature, 72±4° F., and relative humidity ofapproximately 25%. After an initial acclimation period with a 12-hourlight/dark cycle, animals are housed in a 24-hour dark environmentovernight before start of the in vivo phase of the study. Animals havefree access to feed and drinking water and are checked for generalhealth and well-being prior to use in the study. Body weights aredetermined for a representative sample of mice prior to initiation ofdosing. The average weight determined from this sampling is used toestablish the dose for all mice in the study.

Each test compound is dissolved in the control solvent (EtOH), anddiluted 1:10 (vol/vol) in corn oil (Crisco Pure Corn Oil, J.M. SmuckerCompany, Orrville, Ohio) to the desired dose (mg/kg) in the desiredvolume (0.1 mL/animal). The control vehicle is ethanol:corn oil (1:10(vol/vol)). The treatment designations and animal assignments aredescribed in Table 14.

TABLE 14 Dose Group Route Treatment (mg/kg) Animals 1 oral StyrenylCompound 3 5 2 oral Styrenyl Compound 1 5 3 oral Styrenyl Compound 0.3 54 oral Styrenyl Compound 0.1 5 Control oral Vehicle None 5

Animals are dosed once orally by gavage, with the assigned vehiclecontrol or test compounds under red safety light in the dark. The volumeof the administered dose is not to exceed 10 mL/kg.

Four hours after dosing, the mice are exposed to 5000 Lux white lightfor 10 minutes to photobleach their visual pigment. The mice arereturned to the dark and euthanized 2 hours after photobleach byadministering carbon dioxide followed by cervical dislocation. Both eyesof each animal are removed, collected and frozen at −80° C. untilprocessing and analysis. A record of dosing, photobleach, and harvesttimes is maintained.

Eye samples from the in vivo phase of the study are prepared forrhodopsin assays. The rhodopsin assay is conducted essentially asdescribed by Yan et al. (J. Biol. Chem. 279:48189-96 (2004)). All stepsof the rhodopsin assay procedure are performed under dim red light.Typically, two mouse eyes are used per rhodopsin (Rho) measurement.Mouse eyes are enucleated and rinsed with distilled water. The lensesare removed, and the eyes are cut into 3-4 pieces and frozen immediatelyon a dry ice/EtOH bath. Rho is extracted with 0.9 ml of 20 mM BisTrispropane (pH 7.5) containing 10 mM dodecyl-β-maltoside and 5 mM freshlyneutralized NH₂OH. Each sample is homogenized with a Dounce tissuehomogenizer and shaken for 5 min at room temperature (Eppendorf mixer5432). The sample is then centrifuged at 14,000 rpm for 5 min at roomtemperature (Eppendorf Centrifuge 5415R). The supernatant is collected,and the pellet extracted one more time. The combined supernatants arecentrifuged at 50,000 rpm for 10 min (Beckman Optima centrifuge/50.2 Tifixed angle rotor). The supernatant is collected and absorption spectraare recorded before and after exposing the supernatant to a 12-minphotobleach (60-watt incandescent bulb). The concentration of Rho isdetermined by the decrease in absorption at 500 nm when comparingabsorption spectra recordings before and after the photobleach using themolar extinction coefficient (C=42,000 M⁻¹ cm⁻¹). The total opsinprotein level is assumed to be unchanged by the drug treatment and theamount of apo-rhodopsin is calculated based on the reduction ofrhodopsin (Rho) following drug treatment when compared to the vehiclecontrol group. It is expected that the rhodopsin level decreases andopsin/apo-rhodopsin level increases upon administering of one or moresubject compounds to an animal under the conditions disclosed herein.

Animal procedures and data are recorded by group, interval andparameter. All data are recorded at the time observed. In vivo data andinformation are recorded manually. Statistical analysis is conductedusing GraphPad Prism® 4 Software (Irvine, Calif.).

Example 142 Oxygenation of the Retina in an Animal Treated with aStyrenyl Derivative Compound

This example describes determining the effect of a styrenyl derivativecompound on the level of oxygen in the retina of animals after oraldosing of the animals with the compound. The retinal PO2 is measured 6hours after dosing the cat with of a compound (doses range between 0.1to 10 mg/kg) that inhibits isomerization of an all-trans retinyl esteror with vehicle by oral gavage, intra-venous injection, or intra-vitrealinjection. Five cats are dosed with the compound and five cats receivevehicle alone.

The cat is initially given 0.4 mg/kg butorphanol (intramuscularly).Anesthesia is induced with an intravenous injection of 5% pentothalsodium (22 mg/kg) followed by additional pentothal as needed duringsurgery. Intramuscular ketamine (25 mg/kg) is used when the cat isdifficult to handle. Urethane (20%, 200 mg/kg loading dose followed by20-40 mg/kg/h) is used to maintain long-term anesthesia. The cat isparalyzed by an intravenous injection of 2 mL 1% pancuronium bromide(Pavulon; Organon International, Roseland, N.J.) and is artificiallyventilated. Body temperature is monitored by a rectal probe and ismaintained at 39° C. At the end of the experiment, pentobarbital sodiumor saturated KCl solution is injected intravenously to euthanize thecat.

Experimental methods are performed as described previously unless notedotherwise (Linsenmeier et al., J. Gen. Physiol. 99:177-97 (1992); Braunet al., Invest. Ophthalmol. Vis. Sci. 36:523-41 (1995)). The eye isstabilized by attaching the conjunctiva to an eye ring that is part ofthe microelectrode manipulator. Double-barreled oxygenmicroelectrodes(Linsenmeier et al., J. Appl. Physiol. 63:2554-57 (1987)) are insertedinto the eye 6 mm behind the limbus through a guide needle. One barrelrecords a current that is proportional to the oxygen tension of thetissue, whereas the other barrel, filled with 0.9% saline, records localintraretinal ERGs. These electrodes are used to collect both PO2 as afunction of distance across the retina (PO2 profiles) during electrodewithdrawal and the electroretinograms (ERGs) during penetration. Thephotopigment of the eye is photobleached with white light and the oxygenprofile is collected in the retina during dark adaptation. Themicroelectrode penetrates all the way to the choroid, signaled by thetransepithelial potential (TEP) when the electrode crossed the retinalpigment epithelium (RPE).

A one-dimensional, three layer diffusion model (Linsenmeier et al., J.Gen. Physiol. 99:177-97 (1992); Haugh et al., Ann. Biomed. Eng. 18:19-36(1990)) is fitted to the part of the oxygen profile in the avascularregion of the retina to quantify photoreceptor oxygen consumption (Qav).The model assumes that only the layer corresponding to the inner segmentconsumes oxygen. This layer is the location of all the photoreceptormitochondria, with the exception of those in the synapses. Diffusion isassumed to be the only mechanism for oxygen delivery to thephotoreceptors from the choroid and from the retinal circulation, so theouter retina can be treated as a slab of tissue through which oxygendiffuses. The model equations for PO2 as a function of distance from thechoroid has been described by Wang et al., Invest. Ophthalmol. Vis. Sci.48:1335-41 (2007).

Animal procedures and data are recorded by animal, interval andparameter. All data are recorded at the time observed. Parametersderived from all profiles obtained under a given condition in each catwere averaged, and the results are reported as the mean±SD across cats.Paired t-tests are used to test for significant differences between anytwo conditions. The difference is considered significant if P<0.05. Itis expected that the steady state oxygen concentration increases uponadministering of one or more subject compounds to an animal under theconditions disclosed herein.

Example 143 Effect of a Strenyl Compound on Retinal Function inOxygen-Induced Retinopathy

This example describes the effect of a styrenyl compound on rod functionin the retinal. Oxygen-induced retinopathy is induced in 4 groups (sixanimals each) of Sprague-Dawley rat pups by exposing the pups toalternating periods of 50% and 10% oxygen beginning on the day of birth(Day 0) to Day 14. The light cycle is 12 hour light (10-30 lux) and 12hours dark. The light to dark transition coincides with each oxygenalternation. Beginning at Day 7 and continuing for 15 days thereafter,within one hour of the light-to-dark transition, two of the four groupsare administered 6 mg/kg of styrenyl compound intraperitoneally. Onlyvehicle is administered to the other two groups. When marked retinalvascular abnormality is generally observed, at Day 20-22,electroretinograms are recorded and receptor and post-receptor functionare evaluated. The treatment effects are analyzed by ANOVA. (See, e.g.,Liu et al., Invest. Ophthalmol. Vis. Sci. 47:5447-52 (2006); Akula etal., Invest. Ophthalmol. Vis. Sci. 48:4351-59 (2007); Liu et al.,Invest. Ophthalmol. Vis. Sci. 47:2639-47 (2006)). It is expected thatone or more subject compounds reduces oxygen-induced retinopathy whenadministered into a subject.

When ranges are used herein for physical properties, such as molecularweight, or chemical properties, such as chemical formulae, allcombinations and subcombinations of ranges and specific embodimentstherein are intended to be included.

The various embodiments described herein can be combined to providefurther embodiments. All U.S. patents, U.S. patent applicationpublications, U.S. patent applications, foreign patents, foreign patentapplications, and non-patent publications referred to in thisspecification and/or listed in the Application Data Sheet, areincorporated herein by reference, in their entireties.

From the foregoing it will be appreciated that, although specificembodiments have been described herein for purposes of illustration,various modifications may be made. Those skilled in the art willrecognize, or be able to ascertain, using no more than routineexperimentation, many equivalents to the specific embodiments describedherein. Such equivalents are intended to be encompassed by the followingclaims. In general, in the following claims, the terms used should notbe construed to limit the claims to the specific embodiments disclosedin the specification and the claims, but should be construed to includeall possible embodiments along with the full scope of equivalents towhich such claims are entitled. Accordingly, the claims are not limitedby the disclosure.

While preferred embodiments of the present invention have been shown anddescribed herein, it will be obvious to those skilled in the art thatsuch embodiments are provided by way of example only. Numerousvariations, changes, and substitutions will now occur to those skilledin the art without departing from the invention. It should be understoodthat various alternatives to the embodiments of the invention describedherein may be employed in practicing the invention. It is intended thatthe following claims define the scope of the invention and that methodsand structures within the scope of these claims and their equivalents becovered thereby.

What is claimed is:
 1. A method of modulating chromophore flux in aretinoid cycle comprising introducing into a subject a non-retinoidaromatic compound that inhibits 11-cis-retinol production with an IC₅₀of about 0.1 micromolar or less when assayed in vitro, the assayconsisting of a homogenate of HEK293 cell clone expressing recombinanthuman RPE65 and LRAT as the source of visual enzyme, exogenousall-trans-retinol in the amount of about 20 μM, recombinant human CRALBPin the amount of about 80 μg/mL, about 10 mM pH 7.2 phosphate buffer,about 0.5% BSA and about 1 mM NaPPi, and wherein the amount of assayreaction product 11-cis-retinol being determined by HPLC analysisfollowing heptane extraction of the assay reaction mixture and whereinthe non-retinoid aromatic compound consists of a benzene core that issubstituted with a first substituent, a second substituent, and anoptional third substituent, wherein the first and second substituentsare attached to the benzene core in a meta-substitution configuration,wherein the first substituent is a group selected from —CH(OH)CH(R)CHNH₂wherein R is H, CH₃, or —OH and the second substituent is a substitutedalkenyl, and wherein the optional third substituent is a halogen.
 2. Themethod of claim 1, wherein the R is H.
 3. The method of claim 1, whereinthe substituted alkenyl is a 1,2-disubstituted alken-1-yl moiety.
 4. Themethod of claim 3, wherein the 1,2-disubstituted alken-1-yl is a2-(cycloalkyl)-1-ethenyl moiety.